KR20100028526A - Microfluidic and nanofluidic devices, systems, and applications - Google Patents

Microfluidic and nanofluidic devices, systems, and applications Download PDF

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Publication number
KR20100028526A
KR20100028526A KR1020097018397A KR20097018397A KR20100028526A KR 20100028526 A KR20100028526 A KR 20100028526A KR 1020097018397 A KR1020097018397 A KR 1020097018397A KR 20097018397 A KR20097018397 A KR 20097018397A KR 20100028526 A KR20100028526 A KR 20100028526A
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South Korea
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microfluidic
microfluidic device
valve
channel
sample
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KR1020097018397A
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Korean (ko)
Inventor
윌리엄 디 닐슨
누옌 마이클 반
마티어스 방보
알렌 알 보론케이
율류 아이 블라가
스테반 조바노비치
조안 혼
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마이크로칩 바이오테크놀로지스, 인크.
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Priority to US60/899,630 priority
Application filed by 마이크로칩 바이오테크놀로지스, 인크. filed Critical 마이크로칩 바이오테크놀로지스, 인크.
Publication of KR20100028526A publication Critical patent/KR20100028526A/en

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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844Nucleic acid amplification reactions
    • C12Q1/686Polymerase chain reaction [PCR]
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502715Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/50273Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the means or forces applied to move the fluids
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00Heating or cooling apparatus; Heat insulating devices
    • B01L7/52Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
    • B01L7/525Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples with physical movement of samples between temperature zones
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B82NANOTECHNOLOGY
    • B82YSPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y30/00Nanotechnology for materials or surface science, e.g. nanocomposites
    • FMECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
    • F16ENGINEERING ELEMENTS AND UNITS; GENERAL MEASURES FOR PRODUCING AND MAINTAINING EFFECTIVE FUNCTIONING OF MACHINES OR INSTALLATIONS; THERMAL INSULATION IN GENERAL
    • F16KVALVES; TAPS; COCKS; ACTUATING-FLOATS; DEVICES FOR VENTING OR AERATING
    • F16K99/00Subject matter not provided for in other groups of this subclass
    • F16K99/0001Microvalves
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/02Adapting objects or devices to another
    • B01L2200/028Modular arrangements
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06Fluid handling related problems
    • B01L2200/0647Handling flowable solids, e.g. microscopic beads, cells, particles
    • B01L2200/0668Trapping microscopic beads
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/10Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/08Geometry, shape and general structure
    • B01L2300/0809Geometry, shape and general structure rectangular shaped
    • B01L2300/0816Cards, e.g. flat sample carriers usually with flow in two horizontal directions
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/08Geometry, shape and general structure
    • B01L2300/0861Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/0867Multiple inlets and one sample wells, e.g. mixing, dilution
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/08Geometry, shape and general structure
    • B01L2300/0861Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/0874Three dimensional network
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/08Geometry, shape and general structure
    • B01L2300/0861Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/088Channel loops
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/08Geometry, shape and general structure
    • B01L2300/0887Laminated structure
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/08Geometry, shape and general structure
    • B01L2300/0896Nanoscaled
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/18Means for temperature control
    • B01L2300/1805Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
    • B01L2300/1827Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks using resistive heater
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00Moving or stopping fluids
    • B01L2400/04Moving fluids with specific forces or mechanical means
    • B01L2400/0475Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0481Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure squeezing of channels or chambers
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00Moving or stopping fluids
    • B01L2400/06Valves, specific forms thereof
    • B01L2400/0633Valves, specific forms thereof with moving parts
    • B01L2400/0655Valves, specific forms thereof with moving parts pinch valves
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502761Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00Heating or cooling apparatus; Heat insulating devices
    • B01L7/52Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B33ADDITIVE MANUFACTURING TECHNOLOGY
    • B33YADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
    • B33Y80/00Products made by additive manufacturing

Abstract

The present invention discloses the integration of programmable microfluidic circuits for processing biochemical and chemical reactions and for achieving practical use for integrating these reactions. In some embodiments, a workflow or chemical workflow for a biochemical reaction is combined. A microvalve, such as a programmable microfluidic circuit, having a Y valve, and a flow flow are disclosed. In some embodiments, the microvalve of the present invention is used for fluid mixing, which may be part of an integrated process. This process includes mixing a sample, moving the reaction to a reservoir or edge for the modular microfluid, using a capture area, and injecting it into an assay device on a separate device. In some embodiments, bead trapping by varying the cross-sectional area of the channel in the microvalve, or forming and overlap forming designs, is used. Further movement of the sample between temperature zones, using fixed temperature and movement of the sample by the micropump, is further disclosed.

Description

Microfluidic and Nanofluidic Devices, Systems and Applications {MICROFLUIDIC AND NANOFLUIDIC DEVICES, SYSTEMS, AND APPLICATIONS}

Cross-reference

This application claims the priority of US Provisional Application No. 60 / 899,630, entitled "Microfluidic and Nanofluidic Devices, Systems, and Applications," filed February 5, 2007, which is incorporated herein by reference in its entirety. do.

Technology for Government Supported Research

The present invention relates to project number W911SR-04-P-0047 awarded by the US Department of Defense, approval number 5RO1HG003583 awarded by NIH, contract number NBCHC050133 awarded by HSARPA, order number TTA-1-0014 (according to HSARPA). US Government assistance under one or more of Agreement Number W81XWH-04-9-0012. The US government has certain rights in the invention.

A variety of different designs for the purpose of reducing sample volume requirements in bioanalytical methods, integrating into multiple stages of automated processes, integrating sample preparation and analysis, and linking samples and procedures to the full volume world. Microfluidic devices have often been developed in the past.

In the absence of criteria to control external dimensional form factors, the nature of upstream and downstream external interfaces, and the length, cross-sectional shape, and diameter of internal microfluidic paths, such as microfluidic devices, are often mutual and existing upstream purification and downstream analysis. It turns out that it does not match the device.

Despite advances in micromachining that allow for analysis on microliter, even nanoliter or picoliter scales, many biological and environmental samples are first obtained in much larger volumes than the scale of conventional microfluidic analyzers.

Modular microfluidic technology can combine moving samples on a microchip. While the interest in microfluidics has been to integrate multiple functions into a single device, this goal has been achieved through an alternative approach that enables modular integration of functions across multiple devices. This modular concept is based on a technique that allows capillary arrays to be connected by simply plugging two connectors together (US Pat. No. 6,551,839; US Patent Application No. 11 / 229,065; US Patent 6,190,616; US Patent 6,423,536). US Patent Application No. 09 / 770,412; US Patent No. 6,870,185; US Patent Application No. 10 / 125,045; US Patent Application No. 10 / 540,658; US Patent Application No. 10 / 750,533; US Patent Application No. 11 / 138,018; All of which are incorporated herein by reference in their entirety). In addition to creating connectors, the interfaces between the arrays can also create true zero dead-volume valves and routers. The present disclosure provides guidance on how to connect and cut microchips containing fluid circuits to other microchips or capillary arrays, and how to develop new microchip designs that insert two, three or more microchips. New features include designing fractions, fractionation, new applications and instrument manipulations. This new technique is called modular nanofluidics and the use of microchips in modular nanofluids is referred to as "modular microfluidic microchips" or "modular microchips."

Modular microfluidic microchips have a number of applications in life sciences and medicine. Modular microchips may perform a single function or a function of a logically clustered group. More complex processes occur by docking two microdevices and transferring processed samples. For example, one microdevice can perform PCR amplification and cleanup of a series of target DANs, and the second microdevice can perform a cycle sequencing reaction and perform DNA sequencing. Can be connected to a third device. Similarly, for proteomics, one device may perform separation of the first dimension and the second device may perform second orthogonal separation. The ability to connect devices in a "plug-and-play" manner allows the process to proceed independently at different speeds, cycle times or throughput. A set of modular microchips can be manufactured and incubated in a hotel. Once one step of the sample preparation process is complete, the modular microchip for the next step can be docked, loaded and moved to the hotel for incubation of the next step if necessary. Once sample preparation is complete, the modular microchip can be connected to a high throughput CAE microchip, mass spectrometer, flow cytometer or other analytical device.

The modular approach allows macroscale devices to manufacture and analyze nanoscale samples such as automation and robotic operations. The positional accuracy required for modular operation is good within the existing capabilities of current robotic systems and is much less than that required in the microelectronics industry. Stepped and stepped motors are currently capable of positioning below 1 μm, and positioning above 5 μm is quite routine. Modular microchips can affect a wide range of automation capabilities that directly accelerate the development and deployment of nanoscale devices.

Modular microfluidic approaches can combine microchip-based technologies with automated systems to fully automate nanoscale sample preparation methods and form a technology platform for linking them to many types of analyses. In the present invention, examples of how the present invention applies to the development of DNA sequence sample preparation and analysis, AFLP analysis, PCR analysis, MLVA analysis, cycle sequencing, DNA fragment analysis for genotype and fragment analysis for DNA sequencing are provided. do.

In the present invention, guidance is provided on how to connect microscale and nanoscale devices and how to perform sample preparation and analysis on a microchip. The inventors also teach methods of connecting different microchips to perform specialized functions, including microchip valves, reactors, fluid movement, mixing, different biochemical functions, and analytical methods.

Summary of the Invention

In one aspect, the present invention provides a microfluidic device comprising a microfluidic layer, an actuation layer and an elastomeric layer sandwiched therebetween, the microfluidic layer being assembled at a nexus. Three or more microfluidic channels separated by discontinuities that impede fluid flow in the three or more microfluidic channels, the drive layer comprising one or more drive channels that open into a valve chamber disposed opposite the nexus; The diaphragm valve is formed by the displacement of the elastomeric membrane adjusting the fluid flow across the three or more microfluidic channels. In one embodiment, the elastomeric membrane simultaneously regulates fluid flow across the three or more microfluidic channels. In another embodiment, the elastomeric membrane disrupts fluid flow across the three or more microfluidic channels. In another embodiment, the elastomeric membrane incompletely inhibits fluid flow across the three or more microfluidic channels. In another embodiment, the diaphragm valve further comprises a vias layer. In another embodiment, applying pressure or vacuum to the one or more drive channels regulates the fluid flow across the elastomeric membrane to form three or more channel valves. In another embodiment, the membrane naturally closes the valve, and applying a vacuum to the membrane deflects the membrane from the valve seat to open the valve. In another embodiment, the microfluidic layer comprises a surface facing the membrane, the surface comprising a groove forming a microfluidic channel when pressed against the membrane. In another embodiment, the microfluidic layer includes internal microfluidic channels that intersect in holes in the layer that are open to the nexus. In another embodiment, the drive layer includes a surface that resists the membrane, which surface includes a groove that forms a drive channel when pressed against the membrane. In another embodiment, the drive layer includes an internal drive channel that opens to the valve chamber. In another embodiment, the three or more microfluidic channels gather in a Y-shaped nexus. In another embodiment, the plurality of said diaphragm valves are driven by a single drive channel.

In another aspect, the present invention provides a microfluidic device comprising a microfluidic layer, a drive layer and an elastomeric layer sandwiched therebetween, wherein the microfluidic layer is assembled at a nexus and between but between the first and second channels. A first and second microfluidic channels separated by discontinuities that impede fluid flow that does not follow the channel, wherein the drive layer includes one or more drive channels that open into a valve chamber disposed opposite the nexus, The diaphragm valve is formed by the displacement of the elastomeric membrane adjusting the fluid flow across the three or more microfluidic channels. In one embodiment, the fluid flow in the second microfluidic channel is not regulated by the elastomeric membrane. In another embodiment, the elastomeric membrane disrupts fluid flow across one or more microfluidic channels. In another embodiment, the elastomeric membrane incompletely inhibits fluid flow across one or more microfluidic channels. In another embodiment, the diaphragm valve further comprises a valve layer. In another embodiment, applying pressure or vacuum to the one or more drive channels regulates the flow of fluid across the one or more discontinuities. In another embodiment, the two or more microfluidic channels gather in a T-shaped nexus. In another embodiment, the plurality of said diaphragm valves are driven by a single drive channel. In another embodiment, the second channel forms a loop having a predetermined volume and comprises a positive displacement pump of a predetermined volume. In another embodiment, a predetermined volume of liquid is pumped into the first channel of the microstructure device by opening the diaphragm valve and performing a plurality of strokes using a pump.

In another aspect, the invention provides a microfluidic device comprising a microfluidic layer, a drive layer and an elastomeric membrane layer sandwiched therebetween, the microfluidic layer comprising a microfluidic channel, the drive layer being a microfluidic channel. One or more drive channels that open to the valve chamber disposed along the flow path, wherein fluid flows along the channel whether the elastomeric membrane is displaced or not, but the displacement of the elastomeric membrane regulates fluid flow along the channel to form a diaphragm valve. To form. In one embodiment, the elastomeric membrane modulates fluid flow by increasing or decreasing the cross-sectional area of the one or more microchannels. In another embodiment, the diaphragm valve further comprises a bias layer. In another embodiment, applying pressure or vacuum to the one or more drive channels regulates the flow of fluid across the one or more discontinuities. In another embodiment, a microfluidic device is provided that includes a magnet that produces a field in a diaphragm valve. In another embodiment, the magnet is selected from the group consisting of permanent magnets, electromagnets and rare earth magnets. In another embodiment, the path of one or more microchannels forms a star or nested star adjacent the diaphragm valve.

In another aspect, the present invention provides a system comprising a first microfluidic device and a second microfluidic device, the first microfluidic device comprising an inlet, an outlet, a pump configured to pump fluid through a circuit, and a reactor; A first microfluid comprising at least one first functional component selected from a capture zone, a temperature cycling zone, a hot zone, a cold zone, a separation channel, an analysis circuit, a mixer, a bead processing unit and a magnet Wherein the second microfluidic device comprises a plurality of second microfluidic circuits each comprising an inlet, an outlet, and at least one functional component different from the first functional component, wherein the first component and the second component The component is configured to occupy a number of positions, in each of which the outlet of the first circuit is paired with the inlet of one of the second circuits and paired from the first circuit. And a fluid to flow to the second circuit. In one embodiment, the first microfluidic device further comprises an electrode configured to electrically move the fluid, molecule, chemical or particle. In another embodiment, the second microfluidic device further comprises an analysis region. In another embodiment, the second microfluidic device further comprises a capture region. In another embodiment, the second microfluidic device is mobile relative to the first microfluidic device. In another embodiment, the fluid input of the second microfluidic device transfers fluid from the first microfluidic device to a plurality of microfluidic channels. In another embodiment, the fluid output of the first microfluidic device transfers fluid from a plurality of microfluidic channels to the fluid input of the second microfluidic device. In another embodiment, the fluid, molecule, chemical or particle is moved by electrophoresis from the fluid output of the first microfluidic device to the fluid input of the second microfluidic device.

In another aspect, the present invention provides a method of performing a method comprising the steps of: a) performing a first operation on a first sample in a first microfluidic circuit of a first microfluidic device of claim 26; b) engaging the first and second microfluidic devices of claim 26 such that the output of the first circuit is mated with the inlet of the first portion of the second microfluidic circuit; c) moving the first sample from the first circuit to the first portion of the second circuit after operation; d) performing a second operation on a first sample received in a first portion of a second circuit; e) performing a first operation on a second sample in a first microfluidic circuit; f) engaging the first and second microfluidic devices such that the output of the first circuit is paired with the inlet of the next circuit that is different from the second microfluidic circuit; g) moving the second sample from the first circuit to the next of the second circuit after operation; And h) performing a second operation on the first reaction sample in the first portion of the second circuit. In one embodiment, a method is provided that includes repeating steps e) to f) for one or more samples in a first circuit, each sample being moved to a next circuit that is different from the plurality of second circuits. . In another embodiment, the first operation comprises mixing the sample with a reagent. In other embodiments, the first operation is performed in less time than the second operation.

In another aspect, the present invention provides a method of making a microfluidic device, comprising connecting multiple layers to form a plurality of microchannels and diaphragm valves, wherein the plurality of layers are sandwiched together; At least two of the plurality of layers are selected from the group consisting of elastomeric membranes, drive layers, microfluidic layers, valve layers, heat spreaders, bias layers, interface layers and cover layers. In one embodiment, at least one of the plurality of layers is an adhesive layer. In another embodiment, the multiple layers are connected together by an adhesive layer. In other embodiments, the multiple layers are connected together by one or more clamps. In another embodiment, the multiple layers are connected except for the diaphragm valve position by reducing the pressure or temperature at the valve. In another embodiment, the multiple layers are connected except for the diaphragm valve position by selectively placing a coating on the valve. In another embodiment, the coating is removable.

In another aspect, the invention provides a microfluidic channel comprising: a) a microfluidic channel; b) a first temperature zone disposed along a channel having a temperature above ambient temperature; c) a second temperature zone disposed along the channel having a temperature below ambient temperature; And d) a volumetric pump disposed along the channel and configured to pump the liquid into the first and second temperature zones. In one embodiment, a microfluidic device is provided that includes a microfluidic loop thermally coupled to one or more temperature zones. In another embodiment, the sample in one or more microfluidic channels is pumped two or more times between two or more different temperature zones. In another embodiment, the sample is separated from the one or more three valve pumps by immiscible fluid. In another embodiment, the one or more diaphragm valves are located in the temperature zone.

In another aspect, the present invention provides a method of treating a microfluidic device comprising heating or cooling a nucleic acid sequence in the microfluidic device of claim 41; And analyzing the nucleic acid sequence. In one embodiment, the assay comprises sequencing, ligation or polymerase chain reaction amplification, transcription, translation, or simultaneous transcription translation. In another embodiment, the nucleic acid is selected from the group consisting of genomic DNA, mitochondrial DNA, mRNA, tRNA, rRNA, siRNA. In another embodiment, the assay comprises ligation of one or more adapters to the nucleic acid sequence. In another embodiment, the adapter comprises a specific nucleic acid sequence identifier. In another embodiment, the specific nucleic acid sequence recognizer is used as an internal quality control metric. In another embodiment, the assay comprises binding of a single nucleic acid sequence ligated to a bead. In another embodiment, a method comprising amplifying said nucleic acid sequence is provided.

In another aspect, the invention provides a microfluidic layer comprising a microfluidic channel; And magnetic field generating means for generating a magnetic field in a tight bend region, the channels comprising at least one tight bend comprising two channel segments connected to each other and oriented at an acute angle. And paramagnetic particles flowing through the tight bend are retarded by a magnetic field. In one embodiment, a microfluidic device is provided that includes a plurality of tight bends.

In another aspect, the invention provides a microfluidic layer comprising a microfluidic channel; And magnetic field generating means for generating a magnetic field in a second region, said channel comprising first, second and third regions, said second region being first and third regions. The larger cross-sectional area and paramagnetic particles flowing through the second region are retarded by the magnetic field.

In another aspect, the present invention provides a method for producing a reaction mixture comprising: mixing a biomolecule and a reagent in a first reaction chamber on a microfluidic device to produce a first reaction mixture; Moving the reaction mixture to a reaction zone in the apparatus and carrying out the reaction to produce a product mixture; Moving the product mixture to a region in the device to capture the product on paramagnetic capture particles within the region; Moving the particles and the trapped product to a capture chamber in the device in a magnetic field to receive the trapped particles and product in the capture chamber; Cleaning the particles in the capture chamber; And moving the particles and the product to a port on the device, wherein the product can be withdrawn from the device. In one embodiment, the biomolecule is selected from nucleic acids (DNA or RNA), proteins, carbohydrates, cells or lipids. In another embodiment, the reaction is a nucleic acid amplification reaction. In other embodiments, the amplification is isothermal. In another embodiment, amplification comprises thermal cycling of the mixture. In another embodiment, a method is provided that includes performing a second reaction on a reaction mixture or product mixture.

In another aspect, the present invention provides a method for carrying out a chemical or biochemical reaction comprising: a) two or more ports, each configured to receive a sample or to remove a sample therefrom, and one or more pumps configured to pump fluid through a circuit; A plurality of microfluidic reaction circuits comprising at least one reaction chamber comprising means and at least one capture chamber comprising means for capturing particles; And b) one or more dispensing ports in fluid communication with a plurality of microfluidic circuits and configured to deliver a sample or reagent fluid to each of the circuits, the plurality of circuits comprising one of the ports in each circuit. It is configured to operate simultaneously on a number of different materials to be delivered to. In one embodiment, the reaction chamber is configured to be heated or cooled by a heat pump. In another embodiment, the capture chamber is disposed in a magnetic field configured to retard the movement of paramagnetic particles in the capture chamber. In another embodiment, the pump is a volumetric pump.

Quote for reference

All publications and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication or patent application was expressly and individually indicated to be incorporated by reference.

The novel features of the invention are described in detail in the claims. The features and advantages of the present invention will be better understood with reference to the following detailed description, which sets forth exemplary embodiments utilizing the principles of the present invention.

1 illustrates the application of a pump on a microchip to move fluid from an input region to an output region.

FIG. 2 shows a Microscale On-chip Valve (MOV) driven by pneumatics.

3 shows a self priming MOV pump on the top panel, a router on the top middle panel, a mixer on the bottom middle panel, and the capture of beads in the channel on the bottom panel.

4 shows an application using five valves to produce a fluid circuit with two pumps on a microchip sharing a common valve.

5 shows a programmable microfluidic circuit that mixes two fluid streams on a microchip and moves them to a reactor.

FIG. 6 shows a programmable microfluidic circuit that mixes two streams and moves the mixed stream to an edge that can be coupled to another process.

7 shows a programmable microfluidic circuit that inputs a sample from the edge, mixes the sample as the stream merges with the microvalve, and moves it to a reactor, which may be a MOW valve.

8 shows a programmable microfluidic circuit that mixes and loads into a reactor connected to the edge.

9 illustrates a programmable microfluidic circuit that can move a sample from two chambers to a reactor and then move the sample or product to the edge for further processing or analysis.

10 shows a programmable microfluidic circuit that moves a sample from a reactor to a location where a second process, such as another biochemical or chemical reaction, can occur.

11 shows two devices performing different functions on each device.

FIG. 12 illustrates a case in which an assay device may be coupled to a sample preparation microchip with modular microfluidic connections.

13 shows a microchip using a modular microfluidic connection having one or both microchips with electrodes.

FIG. 14 illustrates the case where the capture zone is in one device to concentrate, purify or capture the sample in the zone and transfer it to the second device.

15 shows two channels, each with a capture area.

FIG. 16 shows device A with a capture area connected to device B with multiple analysis channels.

17 shows a device having a plurality of electrodes connected to a device having a capture area.

FIG. 18 illustrates a case where the device of FIG. 17 is connected to a device having two or more separate channels.

FIG. 19 is a circuit of one of eight channel circuits on an MBI-026 microchip comprising MOV based mixing, thermal cycling reaction and bead based sample cleaning on the top panel, and a completed glass MBI-026 microchip on the bottom panel. To show.

20 shows the signal and readlength for noise coming from the MBI-026 microchip.

21 shows an automated microchip with microtiter plates.

FIG. 22 illustrates a Sample Capture and Purification Module (SCPM), which is an apparatus that uses immunomagnetic separation (IMS) and pressure induced flow to dispense magnetic beads, and concentrates and purifies bioagent on beads.

FIG. 23 shows an improved method of construction of a MOV microvalve, referred to as a “Y valve”, with three connections.

FIG. 24 shows a MOV flowthrough valve in which two or more flows can be combined.

FIG. 25 contrasts the use of a microvalve not bonded to the left side and an embodiment of connecting the flow valve of FIG. 24 to another channel to eliminate dead volume.

FIG. 26 illustrates the use of microfluidic circuits with features to improve trapping of paramagnetic particles.

FIG. 27 illustrates varying the cross-sectional area of the microfluidic channel to slow the flow and improve trapping of particles by magnetic, optical, and other methods.

FIG. 28 illustrates changing the cross-sectional area of the microfluidic channel by opening the MOV microvalve to slow down the flow and improve particle trapping by magnetic, optical and other methods.

29 illustrates connecting a reservoir or other fluid source to multiple channels using a MOV Y junction.

FIG. 30 illustrates delivery of reagents or samples, using flow valves to minimize the amount of dead volume at the crossover points allowing the channels at the crossover to be isolated from each other when needed.

31 shows a loop comprising a microvalve, such as a MOV micropump.

32 illustrates a programmable microfluidic circuit that incorporates multiple functions into a compact design.

33 illustrates a hybrid plastic microchip.

FIG. 34 illustrates a programmable microfluidic circuit that incorporates multiple functions into a compact design using a star machining design and detector.

35 illustrates the design of a mechanically tightened microchip.

Fig. 36 shows a photograph of the mechanically tightened microchips seen from above on the upper panel and a photograph of the mechanically tightened microchips seen from below on the lower panel.

37 shows a layout of the mechanically tightened microchips.

38 illustrates an assembly of a microchip using double sided tape or adhesive coating material.

39 shows a microchip made of an adhesion layer.

40 shows a close-up of a valve with an adhesion layer.

41 illustrates an adhesive laminated microchip with a heater and a heat spreader inserted.

FIG. 42 illustrates a plastic laminated microchip with normally closed valves that can be adjusted by programmable air pressure.

Figure 43 illustrates a plastic laminated microchip with a normally open valve that can be adjusted by programmable air pressure.

44 shows a four-layer valve.

45 illustrates the top of a plastic microchip and mask design fabricated for bead cleaning.

46 shows a design of a microchip, the upper part of which may be made of plastic or other material, and the lower part of the image showing an acrylic microchip.

47 illustrates the use of a MOV pump to move the sample between the hot and cold zones.

48 illustrates the use of a MOV pump to move a sample between three temperature zones.

49 illustrates moving the sample between the hot and cold zones without passing through the pump.

50 illustrates throughput increase by using multiple samples moving between temperature zones in a multichannel thermal cycler.

FIG. 51 illustrates throughput increase by using multiple samples moving between temperature zones in a multichannel thermal cycler with cleaning channels.

52 shows two temperature cycles in two microvalve.

Figure 53 shows three temperature cycles in three microvalve.

54 shows three temperature shuttle circulations in the microvalve.

55 is a high level diagram of the workflow and process of the NanoBioPrepSuperPyroSequencing (NanoBioPrep SPS ) process.

56 shows a NanoBioPrep SPS ligation module for 42 ligation microchips.

57 shows a 96 channel capillary cassette.

58 shows two programmable microfluidic circuits for ligation.

59 shows a microchip with 96 ligations using a programmable microfluidic circuit.

FIG. 60 shows magnetic beads trapped in a molding capture region.

61 shows a diagram of emPCR amplification.

62 shows a thermal cycler using circulating water.

63 shows a multiple latching microvalve device.

64 illustrates thermal cycling in an emPCR module.

Detailed description of the invention

In one aspect, the present invention provides guidance on the use of programmable microfluidic circuits and devices for processing biochemical or chemical samples. In some embodiments, the microfluidic process is associated with milliliter or centiliter scale input or sample volume. Additional chemical and biochemical processes, and the integration of multiple processes are disclosed. In some embodiments, micromachining of microvalve with different designs is taught.

In certain embodiments, the microfluidic device of the present invention comprises a microfluidic layer, a drive layer and an elastomeric film sandwiched therebetween. The fluid layer includes a fluid channel adapted to flow a liquid. In certain embodiments, the fluid channel is located on the surface of the microfluidic layer in contact with the elastomeric membrane. In this embodiment, open channels, furrows or grooves may be etched into the layer surface. In other embodiments, the channel may be present inside the layer, for example in the form of a tunnel, tube or via. The inner channel can access the layer surface through a hole from the surface to the channel. In one manufacturing method, two or more sublayers may be provided relative to one another such that when the two sublayers are paired, the one or more sublayers comprise grooves that form closed channels. In this embodiment, one of the sublayers includes a hole that opens to the surface at the nexus or port, for example, to allow flow across the valve. The diaphragm valve of the present invention displaces a predetermined volume of liquid. When three are arranged in series, the diaphragm valve may function as a diaphragm pump that functions as a volumetric pump. Modular devices of the present invention may comprise means (e.g., stepped motors) to move the modules relative to each other such that they can engage and mate with fluid channels, for example, crossing different modules in turn. This allows the fast activity to cooperate with the slower activity by moving the faster activity across the slower activity, causing the faster module to deliver samples to each of the circuits in the slower module. The first single sample can be distributed among a plurality of slower channels, or a number of different samples can be delivered to each of the second circuits.

In another aspect, microstructures are used to move fluid on the microchip. In some embodiments, including but not limited to three or more valves (4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 valves) May create a pump to move the fluid. In one embodiment, the valve is a diaphragm valve. Diaphragm valves can be created by driving the deformable structure. In some embodiments, the diaphragm valve comprises a valve seat. In some embodiments, the diaphragm valve can allow or prevent fluid flow in a channel, such as a microchannel. In another embodiment, the diaphragm valve is produced without a valve seat. In some embodiments, the diaphragm valve can change the cross-sectional area of the channel to regulate the flow rate of the fluid through the microchannel. In another embodiment, the diaphragm valve incompletely inhibits fluid flow. That is, the valve allows some fluid to flow through the valve in both the open and closed positions. 1 illustrates the application of a pump on a microchip to move fluid from an input region on an microchip A to an output region.

In another embodiment, microstructures are used to transfer the fluid containing the analyte of interest. In one embodiment, the analyte is a particle. Particles include proteins, peptides, prions, toxins, infectious organisms (including but not limited to bacteria, viruses, fungi), cells (blood cells, white blood cells, NK cells, platelets, skin cells, cheek cells, sperm cells, trophoblasts) , Macrophages, granulocytes and mast cells), nucleic acids (eg, DNA and RNA, including but not limited to mRNA, rRNA, tRNA, siRNA, mitochondrial DNA, chromosomal DNA, genomic DNA and plasmids), cellular components (nucleus, chromosomes, Including but not limited to ribosomes, endosomes, mitochondria, vacuoles, chloroplasts and other cellular fragments, carbohydrates (including but not limited to polysaccharides, cellulose or chitin) and lipids (including, but not limited to, cholesterol, triglycerides) Not included). In other embodiments, microstructures are used to include, but are not limited to, molecules or chemicals [dioxin, PCB, heavy metals, organophosphates, estrogen analogs, rBSTs, drug metabolites, carcinogens or tetratogens]. Move the fluid containing the analyte.

In another aspect, microstructures are used for sample capture and purification, such as microseparation, microvalve, micropump, microrouter, nanofluidic control, and nanoscale biochemistry. In one embodiment, microstructures are used to reduce and automate complex workflows (FIG. 2). In one embodiment, microscale on chip valve (MOV) valves, pumps and routers, and instrument manipulations to operate them include the NanoBioProcessor platform. In other embodiments, the microchip includes, but is not limited to, reactors, capture zones, temperature cycling zones, hot zones, cold zones, separation channels, analysis circuits, mixers, bead processing units, heat spreaders, Peltier devices, and magnets. It includes a microstructure containing the above functional parts. In some embodiments, the capture region will comprise a binding moiety linked to a substrate, including but not limited to an antibody, Fc fragment, Fab fragment, lectin, polysaccharide, receptor ligand, DNA sequence, PNA sequence, siRNA sequence or RNA sequence. Can be. In other embodiments, one or more regions of the microstructures can include beads, such as magnetically reacting beads. In some embodiments, the beads may comprise binding fractions including but not limited to antibodies, Fc fragments, Fab fragments, lectins, polysaccharides, receptor ligands, DNA sequences, PNA sequences, siRNA sequences or RNA sequences. In some embodiments, the magnetic reaction beads have dimensions smaller than 600 nm, such as 590 nm, 580 nm, 570 nm, 560 nm, 550 nm, 540 nm, 530 nm, 520 nm, 510 nm, 500 nm, 490 nm, 480 Nm, 470 nm, 460 nm, 450 nm, 440 nm, 430 nm, 420 nm, 410 nm, 400 nm, 390 nm, 380 nm, 370 nm, 360 nm, 350 nm, 340 nm, 330 nm, 320 nm, 310 nm, 300 nm, 290 nm, 280 nm, 270 nm, 260 nm, 250 nm, 240 nm, 230 nm, 220 nm, 210 nm, 200 nm, 190 nm, 180 nm, 170 nm, 160 nm, 150 nm , 140 nm, 130 nm, 120 nm, 110 nm, 100 nm, 90 nm, 80 nm, 70 nm, 60 nm, 50 nm, 40 nm, 30 nm, 20 nm or 10 nm. In some embodiments, the magnetically reactable beads comprise iron compounds. In one embodiment, the self reacting beads are ferrite beads.

In some embodiments, the magnetic beads have dimensions of 10-1000 nm, 20-800 nm, 30-600 nm, 40-400 nm or 50-200 nm. In some embodiments, the magnetic beads have dimensions greater than 10 nm, greater than 50 nm, greater than 100 nm, greater than 200 nm, greater than 500 nm, greater than 1000 nm or greater than 5000 nm. The magnetic beads can be dry or suspended in liquid. Mixing of a fluid sample with a second liquid medium containing magnetic beads is known in the art, including those described in U.S. Pat. (Unspecified), filed September 15, 2005, entitled "Methods and Systems for Fluid Delivery." Can occur by any means known in the art.

In some embodiments, if an analyte in a sample (eg, the corresponding or non-sugar analyte) is ferromagnetic or has magnetic properties, the analyte may be used in a magnetic field to produce one or more other analytes (eg, the corresponding or non-sugar analyte). Or can be separated or removed from the sample with reduced analyte. For example, the first analyte is coupled to an antibody that specifically binds to the first analyte, wherein the antibody is also coupled to a magnetic bead. When the mixture of analytes comprising the first analyte magnetic bead complex and the second analyte is delivered to the magnetic field, the first analyte magnetic bead complex will be captured as other cells continue to move through the intestine. The magnetic field may then be removed to drain the first analyte.

The magnetic field can be external or internal to the microstructures disclosed herein. An external magnetic field is one whose source is external to the devices of this specification (eg microchips, diaphragm valves, channels, obstructions) contemplated herein. The internal magnetic field is what the source is in within the device contemplated herein. In some embodiments, the magnetic field is generated by a permanent magnet, such as an electromagnet or a rare earth magnet. In some embodiments, the magnet is mobile and can move relative to the microstructures.

In some embodiments, if an analyte to be separated (eg, a corresponding or non-sugar analyte) is not ferromagnetic or has no magnetic properties, magnetic beads may be coupled to the binding moiety that selectively binds such analyte. have. Examples of binding moieties include, but are not limited to, lectins, polypeptides, antibodies, nucleic acids, and the like. In a preferred embodiment, the binding moiety is an antibody or antibody fragment (eg Fab) that selectively binds to the analyte of interest (eg, erythrocytes, cancer cells, sperm cells, nucleus, chromosomes, leukocytes, epithelial cells, bacteria, virus or fungi). , Fc, sfv). Thus, in some embodiments, magnetic beads can be decorated with antibodies (preferably monoclonal antibodies).

The magnetic particles may be coupled to any one or more of the microstructures disclosed herein prior to contacting the sample, or mixed with the sample prior to delivering the sample to the device (s).

In some embodiments, the system herein includes a reservoir containing a reagent (eg, magnetic beads) that can alter the magnetic properties of the analyte captured or not. The reservoir is preferably fluidly coupled to one or more of the microstructures disclosed herein. For example, in some embodiments the magnetic reservoir is coupled to a size-microchannel, and in other embodiments the magnetic reservoir is coupled to the capture region.

The exact nature of the reagent may depend on the nature of the analyte. Exemplary reagents include agents that oxidize or reduce transition metals, reagents that oxidize or reduce hemoglobin, magnetic beads that can bind analytes, or reagents that can chelate, oxidize, or bind iron, or other magnetic materials. Or particles. The reagents change the magnetic properties of the analyte to generate or increase the attractive force to the magnetic field, to generate or increase the repulsive force to the magnetic field, or to remove the magnetic properties so that the analyte is not affected by the magnetic field. can do.

Any magnetic beads that react to the magnetic field can be used in the devices and methods of the present invention. Preferred particles are those having surface chemistry that can be modified chemically or physically, for example by chemical reactions, physical adsorption, entanglement or electrostatic interactions.

In some embodiments, the capture moiety can be bound to the magnetic beads by any means known in the art. Examples are chemical reactions, physical adsorption, entanglement or electrostatic interactions. The capture portion bound to the magnetic beads can vary depending on the nature of the target analyte. Examples of capture moieties include proteins (such as antibodies, avidins and cell-surface receptors), charged or uncharged polymers (such as polypeptides, nucleic acids and synthetic polymerases), hydrophobic or hydrophilic polymers, small molecules (such as biotin, receptor ligands and chelating agents) Carbohydrates and ions, but is not limited thereto. These capture moieties include cells (eg bacteria, pathogens, fetal cells, fetal blood cells, sperm cells, cancer cells and blood cells), organelles (such as the nucleus), viruses, peptides, proteins, carbohydrates, polymers, nucleic acids, supramolecular complexes, other It can be used to specifically bind biological molecules (such as organic or inorganic molecules), small molecules, ions or combinations thereof (chimeras) or fragments.

Once the magnetic properties of the analyte have changed, it can be used to perform separation or enrichment of the analyte on other components of the sample. Separation or incubation may include a positive selection by using a magnetic field to attract a given analyte into the magnetic field, or may use a negative selection to attract a non-sugar analyte. In both cases, individuals of the analyte comprising the desired analyte can be collected for analysis or further processing.

In some embodiments, microstructures are used in analytical methods, wherein one or more molecules, particles, or chemicals are analyzed. In one example, nucleic acids are analyzed. Analyzes include, but are not limited to, ligation or polymerase chain reaction amplification, transcription, translation, simultaneous transcription translation. In another example, the label binding moiety is bound to a target analyte (such as a cell, protein, cell fragment or nucleic acid). Wherein the binding moiety includes, but is not limited to, an antibody, antibody fragment, receptor, receptor ligand, lectin, polysaccharide or nucleic acid. Labels include, but are not limited to, fluorescent labels (FITC, PE, Texas RED, Cyber Green, JOE, FAM, HEX, TAMRA, ROX, Alexa 488, Alexa 532, Alexa 546, Alexa 405 or other fluorescent dyes) , Radiolabels (including but not limited to P 32 , H 3 or C 14 ), fluorescent proteins (including but not limited to GFP, RFP or YFP), quantum dots, gold particles, silver Particles, biotin, beads (including but not limited to magnetic beads or polystyrene beads).

In some embodiments, MOV pumps, valves, and routers are disclosed that enable transport, processing, and analysis of samples. Externally driven pneumatic induction on-chip valves, pumps and routers have an operating volume of 10 nl to 10 μl, 10 nl, 11 nl, 12 nl, 13 nl, 14 nl, 15 nl, 16 nl, 17 nl, 18 nl, 19 nl, 20 nl, 21 nl, 22 nl, 23 nl, 24 nl, 25 nl, 26 nl, 27 nl, 28 nl, 29 nl, 30 nl, 35 nl, 40 nl, 45 nl, 50 nl, 55 nl, 60 nl , 65 nl, 70 nl, 75 nl, 80 nl, 85 nl, 90 nl, 95 nl, 100 nl, 110 nl, 120 nl, 130 nl, 140 nl, 150 nl, 160 nl, 170 nl, 180 nl, 190 nl, 200 nl, 250 nl, 300 nl, 350 nl, 400 nl, 450 nl, 500 nl, 550 nl, 600 nl, 650 nl, 700 nl, 750 nl, 800 nl, 850 nl, 900 nl, 950 nl, 1000 nl, 1.1 μl, 1.2 μl, 1.3 μl, 1.4 μl, 1.5 μl, 1.6 μl, 1.7 μl, 1.8 μl, 1.9 μl, 2.0 μl, 2.5 μl, 3.0 μl, 3.5 μl, 4.0 μl, 4.5 μl, 5.0 μl Fluid flow can be controlled in operating volumes including, but not limited to, 5.5 μl, 6.0 μl, 6.5 μl, 7.0 μl, 7.5 μl, 8.0 μl, 8.5 μl, 9.0 μl, 9.5 μl, 10.0 μl. , WH et al., 2003. Sensors and Actuators B89: 315-323); US Patent Application No. 10 / 750,533; See US patent application Ser. No. 10 / 540,658; All of which are hereby incorporated by reference in their entirety).

In one embodiment, to modify the membrane and drive the valve, the MOV valve and the pump combine two glass microfluidic layers with a polydimethyl siloxane (PDMS) variable membrane layer and pneumatic layer to open and close the valve (FIG. 2). The microfluidic channels etched into the upper glass fluid wafer are discontinuous and lead the vias to the valve seats that are normally closed through the "via wafer" and the microfluidic channel (Figure 2 top panel). When a vacuum is applied to the pneumatic displacement chamber by a conventional scale vacuum and pressure source, a normally closed PDMS membrane is lifted from the valve seat to open the valve (Figure 2 middle panel). The lower panel of Figure 2 shows a top view of a valve of the same size as another panel.

In another embodiment, a self-priming MOV pump (FIG. 3, top) is made by adjusting the operation of three or more valves and can produce flow in either direction. In another embodiment, the router is made from three or more MOV valves (FIG. 3, upper middle panel). In another embodiment, the MOV mixer (FIG. 3, lower middle panel) mixes the sample and reagents at high speed. In further embodiments, the MOV device suitably acts with magnetic beads to pump or trap a set of beads (FIG. 3, bottom panel).

Normally closed MOV valves, pumps and routers are durable, easy to fabricate at low cost, can operate in high density arrays, and have a small use volume. Arrays of MOV valves, pumps and routers can be easily fabricated on microchips such as the NanoBioProcessor microchip. In one embodiment, all of the MOV valves, pumps, and routers on the microchip are Teflon seats, silicone elastomers, polydimethylsiloxanes (PDMS), polyimides, Mylar, Latex, Viton, polycarbonates, acrylics, santaprenes. It can be produced simultaneously in a simple manufacturing process using a single membrane, such as polyurethane or buna. This technology provides the ability to create complex microfluidic circuits and nanofluidic circuits on microchips.

In one aspect, a method of making a microfluidic structure is disclosed. In one embodiment, the structure is made by sandwiching multiple layers of material together to form a complete microfluidic structure. In one embodiment, these layers are joined together using an adhesive (FIGS. 38-41). In another embodiment, the layers are 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 40 One or more clamps, including, but not limited to, 45 or 50 clamps (FIGS. 35 and 36). In another embodiment, the layers are joined together using an adhesive and a clamp or pin. In one embodiment, the multiple layers of material sandwiched together comprise at least three selected from the group consisting of top cover, microfluidic layer, bias, interface, membrane, pneumatic layer, bottom cover, heat spreader, heater, drive layer and valve layer. Layer. In another embodiment, the layer consists of more than one substrate. Substrates that can be used to produce the microvalve include, for example, quartz, glass (such as silicate glass, including but not limited to Pyrex, borofloat, Corning 1737), silicon and plastics [acrylic, polycarbonate, liquid crystal polymers]. , But not limited to, polymethylmethoxyacrylate (PMMA), Zeonor, polyolefin, polypropylene and polythiol]. In other examples, substrates that can be used as membranes include Teflon, silicone elastomers, polydimethylsiloxanes (PDMS), polyimides, Mylar, Latex, Viton, polycarbonate, acrylics, santaprene, polyurethane, and buna, It is not limited to this. In other examples, substrates that can be used as adhesives include, but are not limited to, adhesive coating tapes based on silicone, acrylic, or other materials in double-sided tapes, thin sheets, or thin films.

In some embodiments, the microstructures comprise a plurality of microchannels. In some embodiments, the microchannels are the same in width and depth. In other embodiments, the microchannels differ in width and depth. In one embodiment, it is characterized by having a channel wider than the average size of the analyte of interest in the sample delivered to the microstructure. In other embodiments, the microchannels are the same or larger than the largest analyte (eg, the largest cell) isolated from the sample. For example, in some embodiments, the microchannels in the microstructures are greater than 50 microns, greater than 60 microns, greater than 70 microns, greater than 80 microns, greater than 90 microns, greater than 100 microns, greater than 110 microns, greater than 120 microns, greater than 130 microns It may be greater than 140 microns, greater than 150 microns. In some embodiments, the microchannels are less than 100 microns, less than 90 microns, less than 80 microns, less than 70 microns, less than 60 microns, less than 50 microns, less than 40 microns, less than 30 microns, or less than 20 microns. In some embodiments, the microchannels in the microstructures have a depth greater than 50 microns, greater than 60 microns, greater than 70 microns, greater than 80 microns, greater than 90 microns, greater than 100 microns, greater than 110 microns, greater than 120 microns, greater than 130 microns , Greater than 140 microns, greater than 150 microns. In some embodiments, the microchannels are less than 100 microns, less than 90 microns, less than 80 microns, less than 70 microns, less than 60 microns, less than 50 microns, less than 40 microns, less than 30 microns, or less than 20 microns. In some embodiments, the microchannels have sidewalls that are parallel to each other. In some other embodiments, the microchannels have a top and a bottom parallel to each other. In some other embodiments, the microchannels comprise regions with different cross sectional areas. In some embodiments, the microchannel has a cross section shaped like a cheese wedge, with the pointed end of the wedge facing downstream.

In some embodiments, the microchannels or microstructures are etched as grooves or trenches for the substrate. In some embodiments, the substrate is punched, drilled, cut or etched. In another embodiment, the microchannels are formed in the substrate as tunnels or holes wrapped in the substrate of a single layer of the microchip.

In some embodiments, the microstructures are formed using standard photolithography. For example, photolithography can be used to form an obstacle photoresist pattern on a silicon-on-insulator (SOI) wafer. The SOI wafer consists of a 100 μm thick Si (100) layer on top of a 1 μm thick SiO 2 layer on a 500 μm thick Si (100) wafer. To optimize photoresist adhesion, the SOI wafer may be exposed to hot vapor of hexamethyldisilazane prior to photoresist coating. After spin-coating a UV sensitive photoresist on a wafer, firing at 90 ° C. for 30 minutes, exposing to UV light through a chrome contact mask for 300 seconds, and developing in a developer for 5 minutes, Post-baking at 90 ° C. for 30 minutes. Process parameters may vary depending on the nature and thickness of the photoresist. The pattern of the chromium contact mask is transferred to the photoresist and the shape of the microstructure is determined.

Etching is initiated when forming the same photoresist pattern as the microstructure. SiO 2 may act as a stopper for the etching process. Etching can also be adjusted to stop at a predetermined depth without using a stopper layer. The photoresist pattern is transferred to a 100 μm thick Si layer in a plasma etchant. Multiple dip etching may be used to uniform the microstructures. For example, the substrate is exposed to a fluorine-rich plasma flowing with SF 6 for 15 seconds, then the system is converted to a fluorocarbon-rich plasma flowing only with C 4 F 5 for 10 seconds, and then all surfaces are coated with a protective film. In subsequent etching cycles, aging with ion bombardment removes the polymer preferentially from the horizontal surface and repeats the cycle several times until it reaches, for example, the SiO 2 layer.

To couple the bonding portion to the surface of the obstacle, the substrate may be exposed to an oxygen plasma prior to surface modification to produce a silicon dioxide layer, and the bonding portion may be attached thereto. The substrate is then rinsed twice with distilled deionized water and dried in air. The sample was immersed in a 2% v / v solution of 3-[(2-aminoethyl) amino] propyltrimethoxysilane in freshly prepared water for 30 seconds and further washed with distilled deionized water to remove the silane on the exposed glass. Passivate The substrate is then dried in nitrogen gas and calcined. The substrate is then immersed in a 2.5% v / v solution of glutaraldehyde in phosphate buffered saline at ambient temperature for 1 hour. The substrate is then rinsed again and immersed in a solution of 0.5 mg / ml binding portion, such as anti-CD71, in distilled deionized water for 15 minutes at ambient temperature to couple the binder to the obstruction. The substrate is then rinsed twice in distilled deionized water and immersed in 70% ethanol overnight for sterilization.

There are a number of techniques in addition to the methods described above that can immobilize the bonding moiety on the area of the microstructure and on the device surface. For simplicity and cost consideration, one may simply choose to physically absorb the surface. Another approach may use self-assembled monolayers (such as thiols over gold) functionalized with various bonding moieties. Additional methods may be used depending on the joining portion to be joined and the materials used to manufacture the device. Surface modification methods are known in the art. In addition, certain cells may preferentially bind to the unaltered surface of the material. For example, some cells may preferentially bind to positively charged surfaces, negatively charged surfaces or hydrophobic surfaces, or chemical groups present in certain polymers.

With different materials including but not limited to pyrex, borofloat, Corning 1737, silicon acrylic, polycarbonate, liquid crystalline polymer, polymethylmethoxyacrylate (PMMA), Zeonor, polyolefin, polystyrene, polypropylene and polythiol Microstructure devices can be made. Depending on the choice of material, different fabrication techniques may also be used. High temperature embossing techniques can be used to make the device from plastics such as polystyrene. Obstacles and other required structures can be embossed onto the plastic to form the bottom surface. The top layer can then be bonded to the bottom layer. Injection molding is another approach that can be used to form such a device. Soft lithography may also be used to form the entire chamber from plastic, or may form only a partial microstructure and then bond to the glass substrate to form a closed chamber. Another approach involves using epoxy casting techniques to form obstacles using UV or temperature curable epoxy on a master with a negative replica of the intended structure. Lasers or other types of micromachining approaches may also be used to form the flow chamber. Other suitable polymers that can be used to make the device are polycarbonate, polyethylene and poly (methyl methacrylate). In addition, metals such as steel and nickel may also be used in the manufacture of the devices of the invention, for example by conventional metal machining. Three-dimensional fabrication techniques (such as stereolithography) can be used to fabricate the device in one piece. Other manufacturing methods are known in the art.

Fluid mixing

The ability to mix fluids on microchips and capillaries is disclosed. 4 shows an application using five valves to fabricate a fluid circuit with two pumps on a microchip sharing a common valve. The term "fluid" means, but is not limited to, liquids, gases and other matrices containing simple liquids, mixtures, beads, particles, gels and other materials well known to those skilled in the art, including only one component. The circuit can move the fluid from the two input regions and mix it to deliver the mixture to the output region on Microchip A. Individual fluid streams may be moved by a pump comprising three or more valves, including MOV valves or other valves. The stream may contain samples, reagents, buffers and other components. The valve can produce actuation of the variable structure and can change temperature, pressure. MOV and other microvalve may be used to combine two or more streams (FIG. 3, upper middle panel). In one embodiment, the MOV valve is self-priming and under computer control, which can be guided in both directions, using the same circuit to simply run two covalent pumps to split the sample into two streams. The sample can be moved from the area mark output to the two area mark inputs.

5 shows the mixing of two fluid streams on a microchip, and the movement to the reactor. In some embodiments, the reactor may be located in a MOV valve to improve optical path length, surface to volume ratio, thermal control, and other benefits for detection. In the examples below, additional methods are disclosed for carrying out reactions with microchips having microvalvees such as MOV valves, pumps and routers.

6 shows the mixing of two streams, and the movement of the mixed stream to the edge which can be coupled to another process. Mixing can be performed using a MOV valve.

FIG. 7 illustrates a reactor that may be a MOV valve or chamber formed by inputting a sample from the edge and combining the sample as the streams merge in a microvalve and using a deflection of a membrane, such as an elastomeric membrane such as polydimethylsiloxane (PDMS). It shows moving.

Three or more streams are also combined into the MOV valve (FIG. 3, upper middle panel) and transferred to the reactor (FIG. 8). The stream can be liquid, particles, magnetic beads, nanoparticles, colloids, gases and other materials. MOV valves allow for easy manipulation of fluids as well as magnetic beads, other beads and particles.

Reaction on the Microchip

In one aspect, the sample can be transferred to a reactor (FIG. 8) consisting of etched channels, microvalvees, vials, flow cells, capillaries, reservoirs, or other structures well known to those skilled in the art. MOV valves can be used to form microchips that combine two or more streams to perform different biochemistry. The sample can then be interrogated in the reactor. This may include, but is not limited to, light sensing such as LIF, absorbance, chemiluminescence, surface plasmon resonance.

9 illustrates the use of two peristaltic pumps after moving the sample to the reactor and then further moving the sample or product to the edge for further processing or analysis as possible below. The term "sample" refers to a fluid, a single compound or element, fluid, aerosol, mixture, DNA, RNA, microRNA, protein, lipid, polysaccharide, cell wall, small molecule, and all other biological materials of a biological sample containing the analyte of interest. Includes, but is not limited to, fluids comprising complex samples, such as fluids such as biological or chemical substances, microbeads or particles comprising components. In addition, the sample may be combined with one or more reagents prior to delivery to the microstructures.

FIG. 10 illustrates moving two samples from the edge into the reactor and then to a location where a second process, such as another biochemical or chemical reaction, can occur. This location can also be used to output the sample using a microfluidic device to move the sample to a robot, pipette or other mechanism. Beads, including magnetic beads, can be used to move the sample if necessary.

Integration of two or more processes

In another aspect, one or more microchips extend modular microfluidic docking and undocking to utilize two or more processes of inserting MOV valves, pumps, and routers. FIG. 11 shows one embodiment where device A prepares a sample for later analysis on device B. FIG. In one embodiment, the device may be a real scale device using a microchip or other means for making tubing, plumbing or samples. In another preferred embodiment, device B comprises a separate instrument comprising a capillary, capillary electroporation system or microchip capillary electroporation; Multidimensional gel and capillary electroporation; Mass spectroscopy, multidimensional mass spectroscopy using HPLC, ICP, Raman spectroscopy, detection based on particles, nanoparticles and beads, imaging with fluorescence, IR, optical analysis systems or any other analysis system well known to those skilled in the art have. The stream or fluid may be placed in a reservoir as in FIG. 6 prior to processing on device A, or may come from a side or other shape as shown in FIG. In this way, microchips with sample preparation functions such as reactions, routers, sample cleaning, PCR, cycle sequencing, sandwich analysis, isothermal nucleic acid amplification, isophoresis, isopotential focusing, hybridization, Samples can be prepared by a number of means, including affinity capture, or other methods well known to those skilled in the art. The sample may then be moved to device B, which may be a microchip, mass spectrometer, analytical instrument, or other detector, including a separation device capable of analyzing the prepared sample.

FIG. 12 illustrates a case in which an assay device may be coupled to a sample preparation microchip having modular microfluidic connections. The sample can then be moved from device A to device B by MOV pumps, external pressure pumps, electroosmotic flow, magnetic separation or other methods.

In one embodiment, an electrode can be inserted to move the sample by a potential such as electroporation. 13 shows one configuration. In one embodiment, the analyte is moved in a first direction. In another embodiment, the non-sugar analyte is moved in a second direction. In some embodiments, multiple electrodes can be used. In one example, after the sample is processed in the reactor, the potential can be pumped to an area where the sample can move from device A to assay device B. This may occur if the two devices are able to move to an adjacent position that may include direct physical contact, intimate contact to a small gap, or additional distal contact separated by a connector or interface. One or more electrodes on device B can be used to complete the circuit to create an electric field that transfers the sample or components of the sample to device B. In another embodiment, an additional electrode is present in the downstream device to which device B is attached. In another embodiment, at least one electrode is in a reservoir connected to device B (FIG. 13).

In other embodiments, the sample can also be moved using a MOV micropump, or other means for capturing the area where the capture portion or binding portion can be attached. In some embodiments, capture regions can be used in methods such as hybridization, affinity capture or other methods known to those skilled in the art. In other embodiments, the capture region may comprise non-specific capture means such as hydrophobic or hydrophilic properties, solid phase extraction, metal bonding, adsorption, non-covalent interactions, covalent photoinductive binding, or other means for capturing the corresponding or non-sugar analytes. May be included (FIG. 14). In one embodiment, Microchip A and Device B can be moved together by modular microfluidics. In one embodiment, the capture region can be used as a region for injection into a capillary electroporation (CE) instrument, a microchip with CE, an analytical device included as capillary array electroporation (CAE). Microchip A may then have functions including, but not limited to, capillary array electroporation (CAE), mass spectroscopy, HPLC, PCR, isothermal nucleic acid amplification, isophoresis, isovoltage focusing, hybridization, and affinity capture. Can be moved adjacent to Microchip B. In other embodiments, the capture zone can be used to concentrate or purify the sample prior to moving to a second device capable of performing further sample preparation or analysis.

In one example, two devices, such as microchips, are in contact with or close to each other. 15 shows two channels, each with a capture area. The channel can then move the sample closer to device B. A potential can then be applied to move the sample from the area for injection to device B. Sufficient voltage may be used to "hopping" the sample across the gap. Samples can be metered into device B if necessary. The number of channels can be adjusted so that device A has more channels than device B and can perform repeated injections, or that device A can have fewer channels than device B. In a preferred embodiment, device B is a microchip comprising separate channels for CAE. In one embodiment, device B may, in one embodiment, comprise a separation capillary that may be connected through a modular microfluidic interface. For 2D separation, if the first dimension is equipotential focusing in device A, device B may be moved repeatedly and a portion of the sample in microchip A may be injected into multiple microchannels on microchip B. FIG. 16 shows Device A with a capture region connected to Device B with multiple microchannels that can be used for analyte analysis such as CAE, mass spectroscopy, HPLC.

In one embodiment, the injection of the sample into the microchip uses a "twin T" injector. While the sample travels across the twin T, the twin T defines a small plug between the twin Ts, while separation can occur as the sample is electroporated from the sample reservoir to the waste (Liu S, Ren H, Gao Q, Roach DJ, Loder RT Jr, Armstrong TM, Mao Q, Blaga I, Barker DL, Jovanovich SB.Automated parallel DNA sequencing on multiple channel microchips.Proc Natl Acad Sci US A. 2000 May 9; 97 (10): 5369-74 .)].

In another embodiment, the separation channel is fabricated in one device, while the injector is made on the second device. In some embodiments, the two devices can be docked or undocked.

In one embodiment, the sample is moved to capture regions on individual capillaries, microchips or other devices. 17 shows an apparatus that can be a microchip with a capture area on the right side. The sample may be moved using a first pump by MOV valves, then electric fields, to remove uncaptured analytes and purify the sample. In one embodiment, the microchip with the capture region can then be moved to a separation device as shown in FIG. 18, followed by injection of the complete sample after desorption. Desorption can be carried out by changing temperature, chemical means, photodesorption or other methods well known to those skilled in the art.

Process integration on microchip for sample preparation and cleaning

In one aspect, MOV techniques can be applied to microchip based sample preparation and analysis for a number of areas such as DNA sequencing, biodefence, forensics, proteomics and cell biology. In one embodiment, MOVE techniques can be used to perform automated cycle nucleic acid sequencing and clearance, such as Sanger sequencing. The top panel of FIG. 19 shows one of eight channel circuits on the MBI-026 microchip, including MOV based mixing, thermal cycling reactions and bead based sample cleaning, with the completed glass microchip bottom panel. Is shown. Using on-chip integration cycle sequencing and clearance to prepare Ready-to-Inject ™ sequencing samples, MBI-026 was used to demonstrate read lengths for 1,000 bases of Phred 15 data on a modified MegaBACE DNA analysis instrument. Used (Figure 20). Using the PowerPlex 16 STR reaction on the MBI-026 microchip, MBI demonstrated 16-plex STR PCR amplification for forensics. Many members, MBI-026 Microchip's MOV mixers, pumps and valves can be used for a wide variety of applications.

In one aspect, the operation of the MBI-026 microchip using NanoBioProcessor technology can be automated using one or more robots (FIG. 21). In one embodiment, the robot under software control moves a sample from the microtiter plate to a microchip (eg, MBI-026) included in the interface device using a fixed tip or probe. In one embodiment, the microchip is connected to a device that may include a function for heating and cooling the microchip. The heating device may include a resistive heater attached to the microchip as shown in FIG. 19. In another embodiment, a microchip in which a resistive heater is placed is used, in contact with fluids of different temperatures, using Peltier / thermoelectric heating, using infrared heating, using different temperature air or gas, or other The microchip is heated by means. In some embodiments, the microchip is cooled using air or other gas (such as a coolant), using a Peltier device, in contact with a liquid (such as a coolant), or using other means well known to those skilled in the art.

Interface with bulky samples

In one aspect, a microfluidic device, such as a microchip, is directly or indirectly connected to a device for making a sample for the microfluidic device. Samples can be concentrated and / or purified.

In one embodiment, the environmental aerosol is processed using a fluid to prepare a sample for further microchip sample preparation and analysis. FIG. 22 shows a Sample Capture and Purification Module (SCPM), a device that utilizes and concentrates pressure induced flow and immunomagnetic separation (IMS) for the distribution of magnetic beads and purifies bioagent on beads. In one embodiment, the prepared sample can be eluted from the beads and the eluate can be transferred to a downstream device such as a microchip, or the beads can be moved to a downstream device. In one embodiment, beads may be introduced into the microchip for reduced real time PCR and / or microscale capillary array electroporation (μCAE). In some embodiments, a vibrating mixer can be used with the microchip to vibrate to mix the samples, reagents and beads (FIG. 22). In another embodiment, the driver moves the magnet to trap and discharge magnetic beads (eg paramagnetic beads) in the microstructure.

In one embodiment, the hardware can be operated by software such as DevLink ™ software or embedded software of the MBI. DevLink defines a set of communication and command protocols in standardized automated structures that are simpler, more flexible, and faster to execute than other software development approaches. The DevLink execution framework is based on a core technology that connects multiple operating systems, development languages, and communication protocols. The software driver wraps the individual smart components of the system, greatly reducing the time required to develop traditional new system software. It is easy to integrate the operation of multiple system modules (pumps, valves, temperature controllers, I / O controllers) based on COM or NET. DevLink provides a professional quality software development system for prototyping through product drainage and maintenance.

Y valve

In another aspect, MOV technology can be used to form a valve that routes the fluid exiting from three or more microchannels. In one embodiment, the router may use a microvalve with or without a valve seat. Figure 23 shows a method of construction of a MOV microvalve called "Y valve" with three connections. FIG. 23 shows the fluid shown in black, and the valve seat shown in white. Valve seats can be manufactured by a number of methods well known in the art, including photolithography, embossing, injection molding, and other micromachining methods. The Y valve is in the closed position and the valve seat isolates the three channels that intersect within the valve. When open, the three channels in this figure are able to communicate with each other. The volume of the valve chamber increases upon opening the valve and decreases upon closing. This volume change causes the movement of the liquid to aid in mixing the liquids together in this valve. In the embodiment shown in FIG. 23, there may be two inlet channels and one outlet channel, or one inlet and two outlets, through which liquid enters.

Flow valve

In another aspect, the MOV valve as shown in FIG. 2 can be a flow valve with two or more liquids together. The flow valve of FIG. 24, referred to as the “T valve”, separates the horizontal channel while allowing fluid to pass from top to bottom through the vertical panel in this example, as shown in the left panel. When the valve is opened as shown in the right panel of FIG. 24, the fluid can move from the vertical channel to the horizontal channel, or from the horizontal channel to the vertical channel.

25 shows the difference between the fluid junctions connecting the three channels. On the left side, a standard MOV valve is shown on the left side of the three-way junction. The flow valve is shown on the right. Both conventional MOV valves and flow valves can be used to regulate a portion of the flow or pump, but if the area from the valve to the channel junction is a dead volume, as in a conventional MOV valve, the dead volume in the flow valve is removed.

The number of channels in this type of valve can also vary. The example shown in FIG. 24 shows one channel through the valve and one channel terminating in the valve. In other embodiments, microstructures can be designed that include a plurality of channels passing through a fluid through a valve and / or a plurality of channels terminated at a closed valve.

Microchips and other fluids On the device  Manipulation of magnetic beads

Molding and Nesting Molding capture  Device

In another aspect, magnetic beads are captured while moving through the channel structure. In one embodiment, magnetic bead capture is most effective at the edge of the magnetic field. In one embodiment, the microchannel structure is designed to take advantage of this property by passing the channel several times through the boundary of the magnetic field. In FIG. 26, thick lines represent channel structures and thin lines represent magnets. In this embodiment, the capture efficiency of this system depends on a number of factors, including but not limited to the shear force applied to the beads by the fluid passing through the channel and the force applied by the magnet. In one embodiment, only a limited number of beads may be captured in each channel segment traversing the magnet when the flow is high. When the beads are trapped in the channel, they occupy a fixed amount of space, effectively reducing the channel cross-sectional area and increasing both flow velocity and shear force. If this balance is reached and exceeded, push additional beads along the channel towards the next magnetic boundary. Microchannel designs with multiple magnetic boundary crossings allow more beads to be trapped in the channel compared to designs with a single magnetic boundary crossing.

In some embodiments, the magnet may be an electromagnetic magnet, or a permanent magnet, including but not limited to rare earth magnets, fixed and moving magnets.

In one embodiment, thermal cycling can be performed using a microchannel design with multiple acute angles. In other embodiments, the lengths of structures with multiple acute angles can be used for mixing. When the liquids are together in the microfluidic system, the laminar flow can keep the liquids in a separated state. Structures with multiple acute angles and multiple rotations cause mixing to occur through dispersion. The circuit on the left panel of FIG. 26 can be compressed into the overlap molding configuration shown on the right or a number of other configurations. The overlap molded part takes up less space than the full molded part.

Of beads due to the change of cross-sectional area of microvalve capture

In another aspect, the shape of the capture area can be altered to increase the cross-sectional area, making the capture of the beads more efficient (FIG. 27). Vertical lines represent channels and black circles represent magnets. As the channel flows through the magnetic field, the width or depth of the channel may increase. For some reason, the beads are more efficiently captured by changing the cross-sectional area. Increasing the cross sectional area slows the speed compared to the normal channel cross sectional area for a given flow rate. Slower flow means less force to push the beads out of the capture area. In addition to the slower speed, the additional volume provides the fluid with a passageway to maneuver around the marble captured in this section. Specific shapes can vary greatly. It can also be used with the shaping and overlapping shaping shapes. This cross sectional area increase can also serve a number of different functions, including reaction chambers or thermal circulation chambers.

Beads inside the valve capture

In another aspect, bead capture in the valve (FIG. 28) utilizes the same concept as capture through the use of channel cross-sectional increase from the above example. The microvalve allows the cross-sectional area of the channel to be changed. Upon closure, the cross sectional area of the channel remains uniform. When the valve is open, the cross-sectional area of the channel in the valve increases significantly, and opening the valve creates a space in the microfluidic channel. As the magnetic particles or paramagnetic particles move through the open valve, the flow may be slow and the magnet may trap the particles. When the magnetic force is properly balanced, closing the valve may release paramagnetic particles. Repetitive opening and closing of the valve can be used to remove paramagnetic particles. In order to increase drag, fluids that interact better with the beads may be passed through or the air and liquid in the bolus may be discharged.

Reagent distribution

In another aspect, the interface between the macro world and the microfluidic device is often one of the difficult problems to solve. While microfluidic devices can use only as little volume of reagent as nanoliters or picoliters for analysis, it is usually necessary to connect even larger volumes of reagents to the assay device. For separation devices, only nanoliter or picoliter volumes are injected and processed, while the device may need access to a larger volume of sample to load this smaller volume. It may be advantageous to minimize the total amount of reagent in the channel to minimize the dead volume.

Common Reagent Reservoir

In another aspect, a common reagent reservoir is shown in FIG. 29, where the central reservoir supplies two channels. In this case, only the minimum volume needed to fill the bottom of the reservoir is shared between the two channels, so the total volume needed to feed the two channels is the total of the design with two channels connected to two independent reservoirs. Less than volume In this example, each of the two supply channels is further divided into two channels using a Y valve. The Y valve separated the three channels entering it. Separation reduces contamination causing unwanted flow in the channel which may be caused by evaporation, capillary forces or other sources. In this embodiment, a single non-useable volume needed to cover the bottom of the well is shared between the four channels to reduce the total amount of fluid required for this system. The intersection is used to divide the number of channels or several times the number of channels pulling from a single well, or the valve is substantially unlimited.

This system can use waste in the opposite direction to the collection. In the example shown in FIG. 29, if the flow is directed to the reservoir, four channels may move waste to a single reservoir. Waste storage that needs to be emptied has little benefit. The reservoir may also include an electrode and connect the four channels to a potential or include a vacuum to attract material through the four channels regulated by a Y valve or other valve.

Reagent delivery system

In another aspect, reagents or samples are delivered by using flow valves to minimize the amount of useless volume at the intersections, allowing the channels at the intersections to end from each other, if necessary. The advantage of this arrangement is that all of the reagents in the chain are pumped using a common set of valves. Using a common set of pumping valves reduces the variation in the ratio of mixing reagents that can be induced by deviations in different pumping valves.

Reagent loops

In another aspect, the microstructure comprises three valves in a reagent loop that act as a micropump for circulating fluid in the flow valve, the loop and the fourth valve, allowing additional material to move into or out of the loop ( Figure 31). Multiple flow valves are available. This loop allows the user to cycle through the minimum volume continuously.

This system can also be used to elute the sample. Circulating the eluent along the capture material allows the sample to elute for more time, allowing the sample high concentration area around the capture material to diffuse and allow higher concentrations of elution.

Integrated design

In another aspect, chemical and biological reactions are performed using a system comprising microstructures. In one embodiment, for DNA sequencing, forensics and other uses, a common reagent reservoir is integrated with the bead capture in the valve and reagent delivery system, as in FIG. 32. Three samples can be loaded into the sample reservoir 80. Reagents from common reagent reservoir 70 may then be pumped through check valve 10 and flow valve 20. Pumps can be pumped using, for example, valves 10, 30, 40 and 50 or external pressure with microvalve to regulate flow. When the flow valve 20 is open, a sample from the sample reservoir 80 may approach and pump it into the flow stream through the microvalve 30 and the microvalve 40 and into the reaction chamber 60. . When thermal cycling is performed, the microvalve 40 and 50 can be closed to separate the reaction chamber. The reaction chamber is shown as a channel in FIG. 32, which may also be present in the valve or move between the valve or channel or other fluid component. It can be moved using on-device pumping or off-device pumping, in which the microvalve in a preferred embodiment induces flow in a certain way, which may be an assay (s) It can be programmed to enable the integration of biochemical reactions to produce. The product (s) can be analyzed on the device or can be moved to another device for analysis.

The apparatus shown in FIG. 32 may then perform a second reaction. In one embodiment, the reaction may be a reaction based on beads that undergo a biochemical or chemical reaction to further process the sample. The response can be queried on the device, or moved to another device for analysis.

In other embodiments, the device is programmed to incorporate multiple reaction steps for DNA sequencing applications. In one embodiment, a cyclic sequencing reagent is mixed into a common reagent reservoir 70 which is mixed with a DNA containing sample loaded into the sample reservoir 80 with a sample which is a PCR, plasmid or other nucleic acid amplification product to be sequenced. . The mixture containing the sample and the periodic sequencing reagent can be transferred to the reaction chamber 60 to perform the periodic sequencing reaction using thermal cycling with a programmable fluid using a microvalve. The cycle sequencing product may then be moved to the product reservoir 90 to move the device for further processing, or in preferred embodiments the cycle sequencing product is moved to the reservoir, and beads such as Agencourt SPRI beads Is added to the periodic sequencing product using appropriate chemistry to ensure that the desired periodic sequencing product bound to the beads separates the product from salts and unincorporated dye label terminators or primers that remain in solution. Rather than attaching the periodic sequencing product to the beads, it is apparent to those skilled in the art that the sequencing of the periodic sequencing products remains in solution and the salt and unincorporated dyes can be reversed. The term “bead” is used in the sense including, but not limited to, particles, paramagnetic particles, nanoparticles, gels, gels having affinity capture properties or nonspecific properties. When beads and periodic sequencing products are included in the reservoir 80, the combined mixture can be opened via the microvalve 20 and 30, and microvalve 40 that can include adjacent or fixed or movable magnets. Pump into. As the flow slows down in the open microvalve and the beads are trapped in the magnetic field, beads like paramagnetic SPRI beads are captured. The beads, such as ethanol, may be added to the reservoir and then the beads processed to remove unwanted impurities such as salts and unincorporated dye labeling reactants. The beads can then be pumped to the product reservoir 90. For periodic sequencing, separation can be used to easily analyze the product on individual devices such as CAE or microchips. Different reservoirs can have different configurations, a single sample can be added to reservoir 70, and multiple reagents can be added to reservoir 80 to perform three different reactions on a single sample. It is obvious.

In another embodiment, the device is used for sequencing or synthesis. Laser induced fluorescence, Raman, plasmon resonance or other methods can be used to maintain the beads in a configuration that can be queried by the detector. In this embodiment, the beads can be held in the reaction chamber 32, and a fluorescent labeling reaction can occur so that the reactants can be moved by a micropump past the beads to be read using a detector. The reactants can then be removed by pumping other fluids to clean the beads to remove the reactants using methods such as pyrosequencing, synthetic sequencing, ligation, or other sequencing chemistry. . The next reagent can then be added using a micropump on the device to move the reagent or using a microvalve to adjust the reagent flow to perform the next round of sequencing. The beads can be held by magnetic, light trapping or other energy methods, or by physical limitations such as trapping into a weir structure (US Pat. No. 7,312,611, which is incorporated herein by reference in its entirety). . In one embodiment, the microstructures include an on-chip packed reactor bed design that utilizes one or more weir structures that enable effective exchange or trapping of beads at a reduced level. In one embodiment, each bead may carry out the reaction, or some beads may comprise additional reactants, as currently used in conjunction with pyro sequencing.

In other embodiments, the beads may be retained and some beads may not intentionally perform a reaction to allow for improved detection. Improvements are obtained by optically separating the signal from the beads with the DNA sample queried by physically separating the beads using signals from other beads by physical dilution of the desired signal. This makes it possible to control the signal from other beads, thereby improving the signal for reaction noise and allowing the manipulation of crosstalk in the channel or chamber. For example, if 100 beads of the same size that do not have a DNA sample to be sequenced are added per beads with DNA, there may be one signal for every 100 beads that can improve crosstalk. Other dilutions are possible.

In another embodiment (FIG. 33), a common reagent reservoir 70, a sample, is used to make use of the mold capture in the manufacture of a device capable of cleaning the reaction product using magnetic or other beads or other purification methods after carrying out the reaction. Rearranges the reservoir 80 and the product reservoir 90 and the members of the microvalve 10, 20, 30, and 40. FIG. 33 shows a different design from FIG. 32 that can also perform DNA sequencing. Samples are added to reservoir 180 and cycle sequencing reagents to reservoir 170 and mixed through microvalve 110, 115, 120 and 130 and then chamber 160 for periodic sequencing or other reactions. Can be moved to). The reservoir 185 can be used to add beads to the reaction product that later migrates to the molding capture circuit 140 for magnetic capture. The cleaning can be performed by adding different solutions, for example SPRI chemistry or chemistry based on other beads, to transfer a solution past the trapped beads to remove unwanted products such as salts or unincorporated dyes.

In other embodiments, the integrated design enables the implementation of various processes. It is possible to properly mix the two reagents together, run the reaction on the mixture and clean the product of this reaction. In this system, active fluid pumping occurs between the two reservoirs, so that additional reagents can be supplied to the reaction. This design has the advantage of being very compact and using less space per circuit, which can be packed into the microchip so that some can be diced to improve microchip yield and functionality.

In another embodiment, FIGS. 32 and 33 illustrate coupling of biochemical and chemical reactions as described for chemical reactions such as SPRI clearance and cycle sequencing. It is apparent to those skilled in the art that the use of microvalve on the device can integrate different biochemical and chemical reactions. For forensics, STR reactions such as the PowerPlex 16 to amplify DNA and bead cleaning to purify the reactants can be used to insert short-term repeating (STR) reactions on the device by inserting a programmable microvalve.

In in vitro transcription reactions, translational reactions or simultaneous transcriptional translation reactions can be performed in programmable microfluidic circuits to analyze DNA or RNA, respectively. It can be used to analyze the effects of additional substances added, such as microRNAs, inhibitors, drug recommendations, chemicals or other substances to be analyzed. It can be used as a screen for the effect of added substances on transcription, translation, or co-transcriptional translation reactions. In one embodiment, the simultaneous transcription translation reaction produces an energy harvest product. In another embodiment, a detector 105 is added to monitor the reaction in the programmable microfluidic circuit.

While the designs shown in FIGS. 32 and 33 are shown in a straight line design, it will be apparent to one skilled in the art that other designs, such as radiant designs, may be used (see Roach, D., R. Loder, T. Armstrong, D. Harris, S. Jovanovich and R. Johnston.Robotic microchannel bioanalytical instrument, Sep. 30 2003. US Pat. No. 6,627,446 ";" Roach, D., R. Loder, T. Armstrong, D. Harris, S. Jovanovich and R Johnston.Robotic microchannel bioanalytical instrument, July 20 2004. US Pat. No. 6,764,648 ";" Roach, D., R. Loder, T. Armstrong, D. Harris, S. Jovanovich and R. Johnston. Apparatus and Method for Filling and Cleaning Channels and Inlet Ports in Microchips Used for Biological Analysis, September 7, 2004. US Pat. No. 6,787,111 ", all of which are incorporated herein by reference in their entirety). In one embodiment, the microvalve may perform a reaction using a chemical or biochemical process to process the material to produce product (s) from the sample. The product consists of a chemical or biochemical composition and can generate energy, stored energy, or use a storage pattern to convert energy in a controlled manner.

Incorporation of a chemical or biochemical process into a method of preparing a product or analyzing a component in a reaction, or a substance added to a reaction to analyze some properties of the substance, is disclosed. The term "material" is used without limitation, and in one embodiment may mean a matter or energy when the reaction is analyzed by light generation, including fluorescence, luminescence or other alteration of the electromagnetic spectrum. In one example, the programmable microfluidic circuitry is directed to a nucleic acid by real-time PCR, mass spectroscopy or other means well known to those skilled in the art, or to proteins by reactions where chemical or biochemical transformations catalyzed by the reactants can be analyzed. The reaction queried by a detector, such as a laser induced fluorescence detector, may be performed. In one embodiment, the device may store energy in bulk or in a pattern using chemicals or biochemicals to store energy in a useful manner such as a visual or optical manner. The device may be programmable or manual to change its behavior. It can be used to alter signal generation in a way that alters optical or other properties by chemical or biochemical transformation. It can be used as a sensor for conditions, as a reporter for identification of physical objects, and can convert objects and energy.

The reservoir can be used to connect programmable microfluidic circuits to other upstream or downstream devices. The robot can be used to transfer or move the fluid into the pipette, or connect the modular microfluidic connection directly to another microfluidic device.

Microfluidic Chip Assembly Method-Thermal Bond Membrane

In one aspect, the microfluidic chip (microchip) is dried from two layers of glass or plastic connected by thermal bonding. Some medical test strips are dried using adhesive (typically in the form of a tape) to connect the plastic layer with other features and channels cut into one or more plastic and / or adhesive layers.

In one embodiment, the programmable microfluidic circuit is fabricated from features and associated features that include a MOV microvalve in the chip. (Hereinafter, we use “valve” to mean any of these classes of features.) In the embodiment shown in FIG. 2, two of the bonded glass with etched microfluidic channels and perforations. The stack, a layer of PDMS for the film, and a pneumatic channel are etched using a stack with a layer of glass that provides a driving pressure for the diaphragm. In this structure, PDMS is a suitable magnetic adhesive for glass, but may be locally separated from it at the valve and treated to reduce adhesion within the valve. Three-layer structure is also possible.

In one embodiment, the pneumatically actuated microvalve is made of plastic material. Since the material is changed to a plastic material for the microfluidic and pneumatic layers, the PDMS membrane no longer has sufficient adhesion to use the same composition as glass and new methods have to be developed.

In one embodiment, all plastic chips were assembled using heat and pressure to thermally bond the microfluidic layer, membrane and pneumatic layer simultaneously. Except for the valve region where the lack of pressure prevents the membrane from binding to the microfluidic layer (if time, temperature and pressure are correct), it is necessary to carefully control this process to ensure that the layers are tightly bound. It can be difficult to evenly bond the entire chip and not to couple it to the microvalve.

34 shows an example of a hybrid plastic microchip. In another embodiment of the invention, one or more of several methods are used to thermally bond the membrane to the microfluidic layer. Each method prevents the membrane from binding at the valve by keeping the temperature of the membrane below the binding temperature, allowing the remainder of the membrane to heat up and compress the microfluidic layer. One factor that aids this approach is that the membrane can be quite thin (eg 25 microns), while the diameter of a conventional valve can be 1 to 3 mm. This makes it possible to use an approach in which heat is applied to a heating plate having regions removed in a pattern consistent with the valve pattern. In order to approach the center of the valve from the side, the heat for bonding must travel at least 20 times the distance that must reach the interface between the membrane and the microfluidic layer. This makes it possible to heat the engagement region while keeping the valve region low enough so that the valve region is not engaged. Relevant areas of the valve seat that should not be joined may also be surface modified by coating, including photolithographic masking, subtractive etch, chemical vapor deposition, and other methods well known to those skilled in the art. The associated area of the valve seat, which should not be joined, may also be maintained at different temperatures by local cooling, or the local fluid in each valve receives less heat and does not reach its Tg so that the microfluid and The assembly that heats the membrane layer may have an area removed from the valve.

In one embodiment, (1) using a heated plate or roller with a profile cut such that contact is made with the chip component only in the region where bonding is desired, typically in all regions other than the valve region; (2) A film is attached to the microfluidic layer by using a laser to selectively heat a predetermined region, and by the method consisting of the following (3) and (4). In this approach, it may be desirable for one of the layers to be transparent to the laser wavelength, while the other layer is opaque to the laser wavelength. In this way, laser energy is absorbed at the interface between the materials most desired by the inventors. An additional hard but transparent layer should be used to apply pressure to the interface. Alternatively, a "clearweld" method can be used. In this method, the interface is painted with a special compound designed to absorb laser wavelengths that easily pass through the chip layer, which can all be transparent. (3) Use high power IR sources and masks to hide areas where heating is not desired. Only the exposed areas can be heated. (4) ultrasonic welding. Depending on the materials used, these heating methods can be optionally enhanced by selectively exposing the surface to be bonded (or not bonded) to UV light and / or plasma to change the bonding temperature.

In another embodiment, the pneumatic drive can be dried in an instrument using a microchip, and the microchip is held again against the instrument in a position that enables the drive of the valve membrane. Using this approach in the method of making and attaching the pneumatic layer is flexible. Some embodiments include (1) cutting the pneumatic channel through a double-sided tape (such as by laser cutting or die cutting), attaching the tape to the cover film, and then attaching the assembly to the microfluidic / membrane assembly or Attaching to the microfluidic / membrane assembly first and then adding a cover; (2) Grinding, sculpting or shaping plastic pneumatic pieces so that the channel is not retained. Use liquid adhesive to adhere to the microfluidic / membrane assembly. (3) Crushing or engraving air pressure patterns through the adhesive side of the tape. (4) using mechanical tightening to maintain the microfluid / membrane assembly relative to the pneumatic assembly. In this case, the air pressure may be part of the chip or part of the instrument if necessary. (5) Thermally bonding the pneumatic pieces to the microfluidic / membrane assembly. Note that a low melting point material can be selected for air pressure to prevent damage to the microfluid / membrane assembly.

In one example, a method of making a plastic microfluidic chip is provided. The material can be selected so that the sample is exposed to only one material. If the support plate is at a low temperature, the microfluidic layer can be kept cold during the bonding process while avoiding shrinkage. Choose a method to generate the required heat. Flexibility in selecting materials and attachment methods for the pneumatic layer (s). This is inexpensive. This does not require the development of significant new technologies. This ensures good bonding along the microfluidic channel, even if the pneumatic channel crosses the microfluidic channel. In existing three-layer co-bonding methods, the lack of pressure at these locations has the potential to prevent good bonding and leak channels.

Mechanically joined microchip

In another aspect, a microchip is manufactured by mechanical bonding of a layer that enables the manufacture of a seal between parts that cannot be joined.

In one embodiment (FIG. 35), mechanically tightened chips enable the use of a wide variety of membrane layers without the need to find bonding techniques for various materials. This example includes layers with different characteristic requirements. Both the microfluidic and pneumatic layers require the ability to retain small features, the membrane layer needs to operate as a MOV valve, and the membrane and microfluidic layers must be compatible with chemical or biological analysis. Mechanical tightening allows the construction of such parts without introducing additional materials such as adhesives. The different layers are aligned and mechanically tightened together. 36 shows a photograph of a chip made in this manner.

Join Chips Reduce Alignment Requirements

In this embodiment (FIG. 37), the fine features that need to be aligned are included in the two layers, the pneumatic layer and the microfluidic and valve layer. Features in the microfluidic layer can be combined by multiple layer sealing methods because both are made of the same material. Features such as ridges and channels can be placed on the microfluidic and pneumatic layers to help seal the membrane layer. The pneumatic layer mechanically tightens the membrane against the assembled microfluidic and valve layer. The reduced area that needs to be tightened in this design reduces the tolerance stack and simplifies the manufacture of microchips.

Adhesive Laminated Microfluidic Chips

In another aspect, a stacked microchip is disclosed. MOV microfluidic chips typically include diaphragm valves and related features such as pumps, routers and variable volume chambers (hereinafter we use "valve" to mean any of these classes), and now Use glass for hard layers and PDMS for diaphragm membranes. The glass is thermally bonded to the glass interface, and the original properties of the film provide sufficient adhesion to the glass interface without the adhesive.

For many applications, it may be desirable to replace the glass with plastic to reduce the cost of the microfluidic chip. The PDMS does not have sufficient inherent adhesion to the plastic tested to withstand the pressure used to operate the valve. In addition, the permeability of the PDMS to gas can cause bubbles in the valve that remain closed. These bubbles can be detrimental to the chip's performance.

In some embodiments, adhesives are used to bond layers to make microfluidic chips (US Pat. No. 6,803,019 and US Pat. No. 6,176,962, which are incorporated herein by reference in their entirety).

In one embodiment, the various layers of the microfluidic chip are bonded using an adhesive, and in some embodiments features such as channels are formed in some or all of the adhesive layer. An important difference from other operations in adhesive laminated microfluidic structures is the inclusion of valve features and associated structures. Two examples of embodiments of the concept are shown in FIGS. 38 and 39. In the first embodiment of FIG. 38, the microfluidic layer, the interfacial layer and the pneumatic layer are made of a double sided tape having suitable channels and holes cut through it, for example by laser cutting. The other layer is also made of a thin plastic sheet having features cut through the thin plastic sheet. Thus, multiple layers can be made of double sided tape.

In another embodiment (FIG. 39), the microchip is constructed by etching the channel to the glass wafer and thermally bonding it to the glass using two adhesives assembled through the layers and in a different order. Thus, this arrangement is possible using multiple adhesive layers, which can be double sided tape. The microfluidic layer may be a thermally bonded plastic or may be made of other materials using other molding methods and / or other bonding methods.

In another embodiment, FIG. 40 shows a close up of a valve constructed by adhesive lamination. When pressure is applied to the valve chamber through a channel in the pneumatic layer, the membrane is deformed until it is pressed against two biases, sealing to prevent fluid in one of the microfluidic channels from passing through the valve to the other channel. The vacuum applied to the valve chamber draws the membrane away through the opening, allowing one microfluidic channel to flow into the other.

In another embodiment, the adhesive lamination method increases the flexibility in the design. There is not much restriction on what can be integrated into the design. For example, FIG. 41 shows an adhesive laminate chip with a heater and a heat spreader dried therein. Different embodiments add electrodes, optical components, other temperature regulating components such as heat pipes or heat sinks, sensors, printing markings. Since the curing of the adhesive does not require heating or only slight heating, the adhesive must withstand or be protected from the pressures used in the lamination process, but this type of addition can be made without damaging the adhesive. ,

Adhesive lamination configurations are an inexpensive way to make microfluidic chips. Flexibility is large in the choice of materials. A wide variety of films and double sided tapes are readily available ready-made. The chip can be kept quite thin, which can be useful for thermal control. If thin enough, it may be flexible enough to be stored in roll form. Features in each layer may be laser cut, waterjet cut or die cut as needed. This is all a quick and flexible method, so that rounding and low volume manufacturing can be achieved quickly. Design changes are possible with short procurement time. The technology required to implement the method exists and there are a number of vendors who can supply the assembly. Can be mass produced quickly. Hybrid constructions that produce one or more layers by different techniques are readily performed. If desired, the interfacial adhesive may typically be made relatively thick to form a fully open valve. Such a valve does not require a vacuum source. It can be closed using pressure, and the valve can be opened when the pressure is removed and the membrane returns to its minimum stress position (flat position). Additional functionalities such as insertion of heaters, heat spreaders, optical components and the like can be easily inserted.

Valve array integrated into plastic microfluidic device

In one aspect of the invention, three layers of the same plastic material are laminated to obtain a microfluidic chip. One embodiment is shown in FIG. Coupling parameters for the same plastic material are temperature and pressure. Under appropriate conditions, the membrane will be permanently bonded to both the upper (microfluid) and lower (drive) plates except for the area above the valve chamber, where the plastic membrane moves downward or pressure is applied when a vacuum is applied to the valve chamber. The pressure conditions are different so that they can move up. The pressure in the area with the valve is different because the valve chamber is not in contact with the lower drive layer because it will experience less pressure during lamination or in the press. If a constant temperature is applied to the entire device or assembly, the pressure will be less for the material in contact in the valve to prevent the bond if the temperature is adjusted such that the bond occurs only when the pressure is transferred to the membrane. This is usually a closed valve. In other embodiments, normally open valves can be produced by applying a higher pressure in the microfluidic channel than in the valve chamber during lamination. In FIG. 43 a shrinked film will be obtained after device cooling. Only pressure is required to drive this valve. Pressure is available without electrical energy, and this valve can be important for portable instruments. For these three layer structures, the air channels cannot traverse more than two microfluidic channels, and vice versa, because the membrane will not bond under the channel (no pressure here). This can be a hindrance to organizing valves in multichannel devices.

In other embodiments, intersecting air and microfluidic channels may be used with the four layer structure. 44 shows a cross section of this structure. This valve is normally closed and requires pressure and vacuum to actuate. Normally open versions can also be manufactured which require only pressure for driving.

In another embodiment, a patternable material that can be selectively deposited on the valve seat and then removed after bonding, such as a photoresist such as Shipley 1818, screen prints of water soluble polymers, UV activated polyacrylamides that match the bonding temperature (ReproGel Photoactive compounds such as GE Healthcare). Alternative embodiments modify the surface properties of some regions by, for example, UV activating a plastic, such as polycarbonate, in the region in need of selective bonding to alter the melting temperature of the modified region. In another embodiment, a liquid such as a viscous liquid comprising acetonitrile, preferentially polystyrene glycol and propanediol, is passed through to prevent binding of the membrane to the valve seat.

Linear array of valves for cleaning beads in plastic microchips

In another aspect, a linear array of valves was designed for the beads cleaning protocol. For example, FIG. 45 shows a mask design on the top panel and a picture of an acrylic three layer chip on the bottom panel. The device was made from two 1.5 mm thick acrylic plates and 0.04 mm acrylic film. Microfluidic channels (blue) and drive channels (red) are milled (0.25 mm wide and 0.1 mm deep) in each one of the 1.5 mm plates, respectively. Drill access holes in both acrylic plates. The special channel for junction temperature control is cut into one plate. The acrylic sandwich was aligned and placed in a polarizing chuck covered with a glass plate and placed in a press with a heating plate. Lamination was carried out at 100 ° C. and 1 ton pressure. Chips were tested for 3 days of continuous pumping. No pump failure was found.

In Figure 46, the design and photograph of a three layer acrylic chip for forensic protocol is shown. The chip will mix the sample with the bead solution, use the bead sample concentrate in the large valve, mix the washed sample with the PCR reagent, and perform PCR amplification and post-PCR cleaning. The chip dimension is 30 mm x 30 mm. The channel is 0.25 mm wide and 0.15 mm deep. The amplification channel can accommodate about 1.5 μl. The microchip works as follows. Pneumatic line 211 drives the microvalve (210, 220). Pneumatic line 212 drives microvalve 215. Pneumatic line 213 drives microvalve 250, 255. Pneumatic line 214 drives microvalve 276. Pneumatic line 216 drives microvalve 240. Pneumatic line 217 drives microvalve 230. Pneumatic line 219 drives microvalve 246. Samples containing material attached to paramagnetic beads are loaded into reservoir 280 connected to microvalve 220. The sample can be moved to a microvalve 240 that is open to pumping by the microvalve 220, 230, 245 and has a magnetic field for trapping beads. The beads are washed using a wash buffer loaded into the reservoir 270 controlled by the microvalve 276 while pumped by the microvalve 276, 230, 245 containing the wash to waste 247. . The reservoir 275 can then be filled with the eluent, and the sample can be purified from the eluted beads with a buffer compatible with the downstream process and then pumped through the microvalve 246. In microvalve 255, the reaction mixture is added from reservoir 270 using microvalve 210, 215, 255 to mix with the eluted sample flowing into reactor 260 as taught. When thermal cycling processes such as PCR or periodic sequencing are performed, valves 250 and 255 can be closed. Following the reaction, the sample is pumped out of reservoir 290 and exported.

These examples were designed for various sample preparation protocols such as sample concentration, thermal cycling (for cycle sequencing and PCR reactions), isothermal reactions, and sample purification. Plastic multilayer stacking can provide low cost, disposable composite chips with valves and pumps. These disposable chips are needed for many applications such as forensics and molecular analysis. Disposable chips can make the instrument much simpler to manufacture without manufacturing and cleaning steps. Normally using an open valve, no vacuum pump or pressure pump will be required. The drive pressure can be provided by the pressurized cartridge and the overall device power requirements can be lowered acceptable.

High speed fluid movement and reaction in the valve

In another aspect, the microstructures move fluid between two different spatial locations. There have been a number of approaches to perform thermal cycling of cycles for PCR, cycle sequencing and other purposes. It can generally be divided into two classes. First, the reaction parts are placed in chambers whose temperature changes over time. Secondly, a constant temperature is maintained at two or more locations, and the reaction components are moved from their location-to-location to achieve temperature equilibrium at each location. We may refer to these as "temporary" and "spatial" cycles, respectively.

In current microfluidic applications, using spatial circulation provides a fluid channel that flows through the temperature zone, which generally leads to a unidirectional reaction fluid through this channel at a constant rate. The flow generally occurs by the application of pressure generated by a device outside the microfluidic structure.

In one embodiment, one or more pumps are used to move the reaction components between zones of different temperatures in each step to achieve thermal cycling. The pump may be external, but in a preferred embodiment, the pump is internal to the microfluidic chip through which the reaction components are thermally circulated. One embodiment is shown in FIG. 47. In this approach, one of our MOV pumps is used to move the sample between the hot and cold zones. For simplicity, this illustration only shows two temperature systems. However, the present invention is not limited to this manner and is easy to extend to the idea of three or more temperatures.

For example, FIG. 48 illustrates a manner of performing three temperature cycles. Here an on-chip pump is used to move the sample through the three temperature zones. In this case, the temperature zone extends over different volumes of channels. While pumping at a constant rate, the sample will be exposed to each temperature for a time proportional to the volume of the channel in each zone. Using on-chip pumping, it may be easy to move each sample instead. In this way, the temperature zone can be made larger and smaller enough to hold the sample. The pump can then be driven for only a short period of time to move the sample from one zone to the next, and the stay at each temperature can be determined by the time between pump runs. This provides the flexibility to adjust the temperature cycling profile without redesigning the channel or heater. Note that the sample can be surrounded by an immiscible fluid to prevent diffusion of the sample. Thus, bolus and microemulsions are driven by this programmable microfluidic circuit and can analyze bolus and microemulsions with a detector to perform real-time reactions or endpoint reactions, such as real-time PCR in bolus or microemulsions. have. For higher throughput systems, it may be desirable to use a pipeline configuration as shown in FIG. 50.

In FIG. 49, an alternative embodiment is shown in which the sample moves between the hot and cold zones without passing through the pump, eliminating interaction with the materials in the pump and preventing the loss of the sample for the dead volume. Again, immiscible fluid can be used for either sample size to help prevent the sample from spreading. It is not necessary to have two complete three chamber pumps as shown herein.

If these features need to be reused, it may be necessary to clean or rinse them before reuse to prevent cross cavity contamination. Additional features may be added for these functions for cleaning the chamber as shown in FIG. 51 for the segment of the reactor.

In another embodiment, the temperature zone is located at the same position as the valve as shown in FIG. 52 for the two temperature cycles and as shown in FIG. 53 for the three temperature cycles. In this embodiment, the bulk of the reaction component can be moved from one zone to another by simultaneously opening the valve while closing the valve comprising it in the target temperature zone. Some of the reaction components may remain in the channel and not reach the intended temperature, which is undesirable. By appropriately sizing the valves and channels, this fraction can be kept small. An advantage of this embodiment is that there is no possibility that the sample moves too much or too little and will not be properly placed in the temperature zone. The reaction within the valve may be present in a three-layer microvalve in which the elastomer or valve membrane is deformed to create a space, or in a four-layer microvalve in which the reaction is carried out through a via.

In FIG. 54, an alternative embodiment for three chamber circulation is shown. In this implementation, the liquid is pumped while rotating. This ensures that the sample does not stay in the channel repeatedly but the channel volume relative to the valve volume is increased.

In some embodiments, there is no need to actually seal the valve in the temperature zone. It is important that only the volume changes as the valve opens and closes. One option to enable this is to leave the valve seats such that channels flow continuously through the thermal circulation zone. This is an advantage during the initial charging of the chip and the movement of the reaction components into the thermal cycling zones. During filling, the minimum required volume of reagents may be used to leave all valves other than one valve closed. If the valve has a seat, it must be open to be fillable. If the chip is charged, it is seen that the reaction component has reached the extreme end of the thermal circulation zone.

In some embodiments, it is desirable to perform thermal cycling in an apparatus having one or more of the following features: rapid temperature transition; Stable and accurate temperature; Small and lightweight package; Disposable reaction vessels; Small reaction volume; Low power requirements; Automatic mixing of samples and reagents; Programmable control of temperature profiles (temperature and time); Configurability for sample size ranges (more than nanoliters); Configurability over a sample number range (1-100 or more per device); Low operating cost; Low pressure requirements for moving the reaction part through the device.

In one embodiment, the microstructured thermal cycler is mobile or compact. In other embodiments, the microstructured thermal cycler is not battery operated. In one embodiment, a fixed zone heating approach is used to keep the power demand unacceptably low and a micropump is used to move the fluid between temperature zones. In other embodiments, microvalves are used to control fluids including analytes, particles, nanoparticles, emulsions, microemulsions, reagents, and combinations thereof.

Reaction in Microchips and Their Uses in Library Construction and Analysis

Conventional Sanger shotgun sequencing has been difficult, expensive and time consuming for in vivo cloning, screening, colony isolation and amplification steps. Large genome centers have automated many of these processes with islands of robotic automation. Next-generation technologies, currently under development or in the early stages of development by companies such as Abiayi, Solexa and 454 Corp., can provide significantly lower cost sequencing by reacting to small ensemble of molecules. At the same time, these techniques (and other techniques such as SuperPyroSequencing and single molecule sequencing) will greatly change the sample preparation requirements and methods.

454 Corp. is currently developing pyro sequencing by performing picoliter reactions on each bead in a "picoliter plate" using shotgun template generation by PCR on emulsion in a reduced form (Margulies M, et. Genome sequencing in microfabricated high-density picolitre reactors.Nature, 2005 Sep 15; 437 (7057): 376-80. Epub 2005 Jul 31.). Each 4 hour operation resulted in about 25 M bases per sequence manipulation with an average read length of about 108. The sample preparation procedure to support each operation is complex with many manual steps. DNA adapters are added to shear the DNA library, single stranded DNA is purified, and DNA is amplified from a single molecule for beads by emulsion PCR (emPCR). Emulsion PCR is capable of producing millions to billions of individual compartments in a single tube using conventional thermal cyclers. Dressman et al. Generated clone PCR amplicons attached to magnetic particles using emulsion PCR [Dressman D, Yan H, Traverso G, Kinzler KW, Vogelstein B. Transforming single DNA molecules into fluorescent magnetic particles for detection and enumeration of genetic variations.Proc Natl Acad Sci US A. 2003 July 22; 100 (15): 8817-22.Epub 2003 Jul 11.) and in Diehl F, Li M, He Y, Kinzler KW, Vogelstein B, Dressman D. BEAMing: single-molecule PCR on microparticles in water-in-oil emulsions.Nat Methods. 2006 Jul; 3 (7): 551-9.), Amplification procedures for complex libraries have been developed (Williams R, Peisajovich SG, Miller OJ, Magdassi S, Tawfik DS, Griffiths AD.Amplication of complex gene libraries by emulsion PCR.Nat Methods. 2006 Jul; 3 (7): 545-5)]. The process of generating a clone library on beads takes about two days using a number of manipulations using specialized equipment, and beads of about 375 bp are prepared. Sample preparation is a major rate limiting step for G20 throughput. It is not possible to generate more than 100-fold increase in bead preparation to sequence the human genome per day without full automation. To extend 454 sample preparation to new sequencing, mammals and other complex genomics require rapid lineage cloning strategies and processing of hundreds of thousands of libraries per day. The same sample preparation can be applied for SOLiD sequencing by ligation.

In one aspect, scaled sample preparation is used to automate library processing into a bead library using microfabricated structures, microfluidics, and magnetic separation in combination with improved emPCR. Large numbers of parallel beads libraries can be constructed to enable automated lineage sequencing approaches. For example, 4,032 individual libraries can be generated every 45 minutes using each monoclonal microsphere having sufficient DNA template for pyro sequencing. In some embodiments, the device may consist of hundreds of thousands of microchannels capable of processing one channel up to millions of individual sequences.

The example describes DNA pyrosequencing, but the DNA analysis method may be a synthetic sequencing such as Solexa, Helicos, ligation reactions such as Agencourt / Applied Biosystem on the sequence, and other reactions such as other DNA analysis methods. .

In addition to generating libraries for DNA sequencing, the same approach can be used for DNA libraries or other DNA analysis, RNA analysis, preparation of samples for proteins, carbohydrates, lipids and other biomolecules. This includes samples prepared by programming in in vitro DNA-RNA-protein systems such as simultaneous transcription translation systems, systems that make modifications to biomolecules, and products from these systems. Similarly, the approach can be extended to chemical libraries and synthesized biomolecular libraries.

The approach can also be used to carry out molecular biological reactions in devices such as microchips.

In the examples below, ligation is used in a number of examples. In addition to the ligation reaction, those skilled in the art can readily perform all biochemical and chemical reactions such as restriction, DNA modification, polymerization, DNA-protein binding and other biochemical reactions as well as addition, removal, cyclization, condensation and other chemical reactions. I can understand.

Motorized beads based Cloning , And based on microbeads In vitro  Emulsion phase PCR  Amplification in Fluid Systems for Library Configuration

In one aspect, beads-based cloning and amplification methods and instrument manipulations can be based on automated instruments to rapidly process multiple sequences. For example, 128,000 input BACs or other libraries can be processed into 250,000,000 beads that can be used for SuperPyroSequencing (SPS) of human-sized genomes daily. 55 illustrates a high level workflow and process. The term “SuperPyroSequencing” is used to describe the reading of beads made with NanoBioPrep SPS where the beads contain a DNA library made from a single copy of the template. In other implementations, different chemistries may be applied to additional workflows for performing real-time PCR in bolus, emulsion and microemulsions. Other biochemistries are possible as described herein. The platform of the NanoBioPrep SPS is an example of combining bead based chemistry for sample preparation for DNA sequencing runs. In some embodiments, gene expression, protein expression, and other implementations can be adapted to bead based reactions and in fluid reactions (US Pat. No. 6,190,616; US Pat. No. 6,423,536; US Appl. No. 09 / 770,412; US Patent No. 6,870,185; US Application No. 10 / 125,045; US Application No. 11 / 229,065; all of which are incorporated herein by reference in their entirety). In some embodiments, the bolus of the sample in immiscible fluid or microemulsion is analyzed as in the SuperPyroSequencing example. This is another example of the present invention that teaches how to couple biochemical and chemical processes and perform multiple reaction steps using a microfluidic device. It will be apparent to those skilled in the art that the modular microfluidic connection may allow for the movement of a sample prepared as described herein.

In one embodiment, the NanoBioPrep SPS can input 4,023 terminal polishing libraries per 45 minutes and 4,023 nanoscale individual ligation of keyed adapters on 42 microchips with 96 channels each within the ligation module. Can be performed. The ligation module then pools the library and joins a single fragment to a single bead. The emPCR module uses emulsion phase PCR (emPCR) to generate beads-based DNA libraries for SuperPyroSequencing.

In one embodiment, to fully automate a microchip portion of the ligation module, a single channel microchip may perform ligation and initiate development of scalable hardware, microchips and software. The 'BeadStorm' device, described below, can resuspend, mix, process, combine and move magnetic and non-magnetic beads. For the emPCR process, hardware that accommodates 100 ml or more of the emPCR reaction and other portions of the workflow is disclosed for a single emPCR, and other implementations may use additional channels that are within the scope of the present invention. Robust, real scale procedures can be used to prepare high quality bead libraries.

In one embodiment, an integrated sample (collectively, such as a bacterial artificial chromosome (BAC), BAC equivalence library, reverse transcript product, single sample, whole genome amplification, environmental samples, including an aerosol collector, for monitoring, including biological defense applications) To prepare beads, plastics, polystyrene, glass or other beads containing cloned individuals of DNA fragments derived from the library, referred to as a library, the NanoBioPrep SuperPyroSequencing (NanoBioPrep SPS ) process (FIG. 55) ligated the adapter and single stranded DNA library And automate library processing to perform emulsion phase PCR (emPCR). The final product, the DNA Bead Library, provides a SuperPyroSequencing reading instrument for collecting complete new mammalian scale genomic sequence determinations per day.

In this example, the NanoBioPrep SPS process inputs 4,032 individual libraries per operation, pre-sheared to individuals gathered at about 1,000 base pairs via end grinding and phosphorylation from 10 ½ 384 well microtiter plates every 45 minutes. do. Thus, the NanoBioPrep SPS can process 128,000 libraries per day, generating 250,000,000 beads. The amount of beads generated will be adjustable. Each bead will contain up to 1,000 bp of amplicons sufficient to generate 60 B bases (human size genome at 10 × coverage per day) of sequence data per day. An example of a NanoBioPrep SPS instrument is the effect of preparing a tip sample to make a shotgun library available for SuperPyroSequencing for new resequencing crystal forms as well as other microbeads and microemulsion processes that can be combined with the biochemical and chemical processes possible in the present invention. Teaches effective and efficient means;

In another embodiment, the NanoBioPrep SPS instrument integrates two modules as shown in FIG. 55. The ligation module enters a fragmented, sized, and end polished library, which (1) binds the specific adapter to the fragment end, (2) recovers the 3 'nick resulting from the ligation reaction, and (3) the second It is denatured to generate a single stranded (SS) DNA library for input into the module, the emulsion PCR (emPCR) module. The emPCR module (1) anneals the SS library on beads using bound primers bound under certain conditions to bind a single DNA molecule to a single bead, and (2) performs emPCR to amplify DNA on the beads, ( 3) Deform double stranded DNA to make glass beads covered with SS amplified library fragments, and (4) Incubate DNA containing beads from null beads by capturing other magnetic beads using annealing primers. The final SS bead library is fed to the SuperPyroSequencing instrument for sequencing and analysis.

NanoBioPrep SPS Ligation  module

In one aspect, integrated gaps to use MOV valves, pumps, and routers to automate the individual ligation of 4,032 different adapter pairs to 4,032 different libraries of abrasive pieces per operation, and then to generate key coded single stranded libraries. An apparatus is provided that includes a large number of mechanisms based on parallel microchips in which recovery and denaturation occur. The NanoBioPrep SPS ligation module may consist of 42 microchips with 96 channels each (FIG. 56). Each channel of the microchip can process a separate library of fragments by ligation of two adapters for each fragment. A single NanoBioPrep SPS microchip is described that adds adapters simultaneously for 96 libraries.

Library loading on microchips

In one embodiment, a library in a microtiter plate, such as a 384 well, can transfer 96 samples to a microchip at a time using an array of 9 mm spacing capillary delivery tubes called capillary cassettes (FIG. 57). Fewer or more capillaries can be used. 58 shows a photograph of a 96 channel capillary cassette used for the reaction. For this NanoBioPrep SPS example, the capillary can be just long enough to be immersed in a microtiter plate well and just wide enough to be filled by capillary action. After filling from the microtiter plate, the transfer capillary cassette is moved by a robot to a 96 input well of a Microchip with 96 ligation circuits in an 8 × 12 format at 9 mm intervals. The capillary cassette is then simply cleaned before reuse to fill the next microchip. We have previously dried capillary cassettes and cleaning stations.

adapter Ligation  reaction

In one embodiment, the microchip or other device can move the sample from the array of capillaries using pressure or vacuum, or electrically induced centrifugation, or other force well known to those skilled in the art. For example, a 96 on-chip MOV pump attached to the input well may pump the library into the microchip as shown in FIGS. 59 and 60. Samples are mixed with 96 individual ligation mixtures in a MOV mixer and pumped into the nano ligation reaction chamber (FIG. 58), where it is incubated for example for 5 minutes. This produces a mixture of adapters attached to the input shear library sample DNA. FIG. 58 shows two ligation reaction circuits, while FIG. 59 shows the mask design for a 96 channel NanoBioPrep SPS .

In one embodiment, each adapter may contain a key, such as six bases, that serves to identify the library after sequencing, provided that each set of 4,032 libraries is achieved by simply distributing the reactor. Separately for each SuperPyroSequencing analysis operation that can be performed). The adapter may comprise overlapping PCR and sequencing priming sites, blunt 3 'ends and 5'overhangs; The adapter of one of each pair may comprise biotin on the 5 'end. The six base sequences generate 4,096 individual keys and 42 microchip devices with 96 channels require 4,032 unique keys, so the remaining 64 keys are available to monitor optimal performance. It can be inserted into each NanoBioPrep SPS operation and used for quality control. After ligation, each ligation fragment is composed of one terminal truncated genomic library DNA and a PCR priming site, a sequencing primer site and six nucleotides long on the key sequence as well as a second end with both PCR and sequencing priming sites. Will contain the key sequence.

In one embodiment, after ligation, 96 uniquely coded libraries are first integrated from the first microchip, then integrated for all 42 microchips, and 20 quality control key sequences are added, with 10 controls in one group. The sequences were difficult to "normal" sequencing, the five control sequences increased difficulty in sequencing, and five control sequences, including all four homopolymer manipulations, were difficult to sequence.

In one embodiment, a reaction chamber such as 800 nl may be used, or other volumes of reaction such as 400 to 200 nl may be performed. The 384 channel microchip can, for example, use 42 microchips to generate a ligation module capable of processing 512,000 libraries per day and can be scaled linearly using additional microchips or instruments.

In one embodiment, pre-binding one of the adapters, for example via biotin-streptavidin binding to paramagnetic beads, results in a beads binding with each adapter at each end to manipulate 4,032 reactions using a magnet after ligation. Fragments can be generated. MOV micropumps routinely interact with beads in microchips and trap beads in channels with magnets (FIG. 61).

Break recovery

In one aspect, after ligation, beads (or libraries) from all microchips in the device can be integrated for beads trapped by a magnetic field in a 5 ml recovery reactor. Bead vortex and magnetic bead driving devices (developed for biological defense applications) used in the sample capture and preparation module operate a device called the BeadStorm ™ device for dispensing, mixing, and trapping magnetic bead suspensions in a temperature controlled chamber. . Gaps at the ends of all ligation adapters will be repaired by Bst DNA polymerase. After gap repair, if the DNA is not yet attached to the beads, prewashed and parallelized streptavidin beads are added and the DNA attached by biotin bound to a set of adapters is recovered. The beads were washed to remove desorbed fragments and adapters, denatured with alkali, and then another set of eight key single stranded QC sequences was added. The ejected single strand was transferred to the emPCR module and discarded magnetic microspheres. The recovery reaction and denaturation step should take about 40 minutes and can be performed in a single batch per operation. The upstream ligation module microchip is regenerated and other manipulations can be initiated while the recovery reaction is being processed.

In one embodiment, the quality of the library can be assessed using an Agilent 2100 or similar device, or the NanoBioPrep SPS instrument automation can create a uniform library that eliminates the need for quality checks at this point. If necessary, a single channel CE component can be added for the ligation module.

NanoBioPrep SPS emPCR  module

In one aspect, the NanoBioPrep SPS emPCR module, as shown in FIG. 61, binds DNA to an optically transparent capture bead to provide for detection of downstream sequencing reactions, to prepare emulsion PCR (emPCR) at or below 100 ml volume. Thermal cycling, denature the resulting amplicons, and isolate beads containing DNA.

DNA capture  Bonding to beads

In one aspect, the SS DNA produced in the ligation module using ligation priming and key sequences can be annealed to other beads or polystyrene containing complementary DNA capture primers. Beads can be made separately from the device. The mold can be annealed by heating the solution to 80 ° C. in a 25 ml MBI Beadstorm processor and then decreasing the temperature while maintaining every 10 ° C. Temperature ramp and hold times can be optimized individually. Another eight key single stranded QCs sequenced can be added at this stage.

Emulsion phase PCR  Amplification

In one aspect, after emulating the emPCR reaction and breaking the emulsion, the beads comprising the amplified double stranded product can be purified. About 30 ml of PCR reaction mixture can scale 42 microchips to a total volume of 100 ml per operation. Since the total time for the emPCR is about 6 hours (this would include 8 45-minute operations), nine parallel emPCR reactors would be required, each of which in this example had a 100 ml emPCR reaction for throughput. Can be processed.

Samples for each emPCR of 4,032 libraries can be flowed into a 100 ml tube operating through an emPCR reactor, and alternatively, emPCR may be performed between two plates. Once the polymerase is thermally activated in the pretreatment chamber, the sample can be transferred to a conventional reaction chamber capable of circulating up to nine simultaneous large volume PCR reactions. The last 12 key QC sequencing beads are added at this step.

In one embodiment, emPCR can be performed first in a typical 96 well microtiter plate, but eight parallel emPCR reactors are designed. For large-capacity heat circulation, in one embodiment three circulation baths with selector valves can produce a thermal circulation mechanism capable of handling large volumes with an excellent ramp. This instrument manipulation was previously dried (FIG. 62) and has been found to enable high speed thermal cycling (due to the optimal heat transfer properties of water) to be well adapted to cycle sequencing and PCR. The software can operate a full scale valve that controls three circulation baths and selects the water and temperature to be circulated. Emulsion chemistry can produce a single dispersed emulsion with sufficient capacity to produce a fragment of 1,000 bases long per microemulsion. In addition to using three water circulation sources, the temperature can be changed by circulating air or liquid at different temperatures. In a simple implementation, three sets of hair dryers set at substantially three different temperatures can be used to quickly change the temperature of a sample. The heated air can be recycled or a fixed temperature can be maintained by operating the heater or hair dryer at a constant temperature.

Single strand DNA  Hatching of beads

In one aspect, after collecting beads with single-stranded DNA, for example, (1) replacing MPC-S with our custom device, and (2) settling or filtering the centrifugation step to change the buffer and wash the beads. And (3) by adjusting the procedure for the density of optical beads rather than Sepharose beads can be hatched using magnetic pull-up by the method of Magulies et al. To collect the emPCR beads from the emulsion, isopropyl alcohol can be added to the PCR amplified emPCR solution, the solution mixed and put through a sieving filter using a syringe pump. After the filter is washed and transferred to the beadstorm, an anneal buffer containing 0.1% Tween is added thereto and the amplified DNA is alkali denatured. After incubation, single stranded DNA containing beads were concentrated by sedimentation, the supernatant was removed and annealing buffer was added. After washing, the magnetic capture beads were added to purify single stranded DNA containing beads by pull-down, the supernatant and the unbound beads were removed, and then the melt was added to break the hybridization. While trapping the magnetic beads, the supernatant containing the hatching beads containing the single stranded template DNA was removed. The NanoBioPrep SPS can then be output to an integrated and key purified library available for SuperPyroSequencing.

42 microchips Ligation  Device and emPCR  Module and Integration

In one aspect, the ligation module can extend from 1 to 42 microchips and can design 42 microchip devices using latching valves as described below. Workflows using a single microchip ligation module and a single channel emPCR module can be used to process a library of cancer cells derived from humans in a non-integrated workflow. To produce automated integrated prototypes that can handle 96 libraries per operation, single ligation module microchips and downstream processes can be integrated with a single channel, flow 100 ml emPCR thermal cycler.

In one embodiment, to complete the NanoBioPrep SPS , 42 microchip ligation modules may be integrated with a 9 channel emPCR module as disclosed herein. Microchip ligation modules can be integrated physically and software using a 1 channel emPCR module in a breadboard. A total of 42 microchip ligation modules can be built and integrated with the 9-channel emPCR module. Fully integrated NanoBioPrep SPS can be optimized and samples can be analyzed on a SuperPyroSequencing device to achieve pyrosequencing, synthetic sequencing, ligation sequencing to achieve a system that can complete new sequencing of humans in one day. Or DNA sequencing by other methods well known to those skilled in the art.

In one embodiment, the ligation module can extend from 1 to 42 microchips. Latching valves can be used to control 42 microchips and begin designing complete system hardware. In one embodiment, a latching microfluidic valve structure is used that is independently controlled using an on-chip pneumatic demultiplexer (FIG. 63). These structures are based on pneumatic MOV valves and usually depend on the nature of the closing. Vacuum or pressure pulses as short as 120 ms are suitable for keeping these latching valves open or closed for several minutes. It has also been found that the on-chip demultiplexer requires only n pneumatic inputs to control 2 (n-1) separate latching valves. A latching valve consisting of both three and four valve circuits has been demonstrated [WH Grover, RHC Ivester, EC Jensen and RA Mathies. 2006. Deveolpment and multiplexed control of latching pneumatic valve using microfluidic logical structure.Lab-on-a -chip.Electronic publication)]. In fact, this technology moves a bank of solenoids over a microchip, reducing cost and complexity.

In one embodiment, the pneumatically controlled NanoBioPrep microchip can be dried using, for example, a latching valve on 42 microchips. The capillary cassette movement mechanism contacts the robotic system with the capillary cassette to load 2,500 valves into four microchip devices.

In one embodiment, the emPCR module can handle 100 ml, for example in a single channel. The high capacity circulating thermal circulator (as shown in FIG. 64) can be dried together with three different independent thermal circulating zones. Nine separate 100 ml emPCR reactors will be needed to share the main thermal cycler that can carry out the same circulation protocol continuously. The pretreatment chamber may facilitate adhesion to polystyrene or other beads and prepare emulsions before adjusting the temperature to meet rapid start requirements. The aftertreatment chamber allows for aftertreatment requirements such as emulsion breakdown and extraction. All three thermal circulation chambers can share bulk circulating water and add water temperatures of 4 ° C. and additional temperatures as needed, which are simply other circulation baths and three real scale COTS valves and solenoids. Engineering surface calculations, research and experiments allow for the surface-to-volume of a reaction, as evidenced by scientific experiments that limit performance and are permitted by the requirement of uniformity of circulation to define the optimum shape for the reactor, e.g., long thick tubes, elliptical tubes or parallel plates. The effect of ratio and depth (mm) can be measured. Separation of these beads carrying amplicons from the null beads can occur in the beadstorm module as a final step before SuperPyroSequencing or other analytical methods. Hardware for executing the device may be integrated under software control.

For example, a single microchip ligation module and a single channel emPCR module can be used to process human cell line libraries in a non-integrated workflow. The control process can be performed manually using GS20 reagents and procedures.

In another embodiment, the NanoBioPrep SPS can be scaled up to fully integrate the ligation module and the emPCR module. A total of 42 microchip ligation modules can be dried by means well known to those skilled in the art. For example, (1) the tower can be built to hold the microchip, (2) the microchip controller can be operated to operate the latching MOV valves, pumps and routers, and (3) the NanoBioPrep robot platform and the first 42 The microchips can be micromachined and tested, and (4) 42 microchip ligation modules can be integrated and tested. At the same time, beadstorm designs can be deployed to handle integrated samples. The emPCR module can be extended to maintain nine operations from the ligation module. Both the 454 GS20 and SuperPyroSequencing can be used to optimize emPCR performance. 42 microchip ligation modules can be plumbed with fully expanded emPCR modules and processes optimized based on the original developed microchip-one channel devices. As described, NanoBioPrep SPS can be used to complete 10% or fully human novel sequencing of the fully human genome.

While preferred embodiments of the invention have been shown and described herein, it will be apparent to those skilled in the art that these embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be used in the practice of the invention. It is intended that the following claims define the scope of the invention and that the methods and structures within such claims and equivalents thereof shall be encompassed by the invention.

Claims (75)

  1. A microfluidic device comprising a microfluidic layer, an actuation layer, and an elastomeric layer sandwiched therebetween,
    The microfluidic layer comprises at least three microfluidic channels assembled at a nexus and separated by discontinuities that impede fluid flow in the at least three microfluidic channels,
    The drive layer includes one or more drive channels that open into a valve chamber disposed opposite the nexus,
    And a diaphragm valve is formed by adjusting the flow of fluid across the three or more microfluidic channels by displacement of the elastomeric membrane.
  2. The microfluidic device of claim 1 wherein the elastomeric membrane simultaneously regulates fluid flow across the three or more microfluidic channels.
  3. The microfluidic device of claim 1 wherein the elastomeric membrane disrupts fluid flow across the three or more microfluidic channels.
  4. The microfluidic device of claim 1 wherein the elastomeric membrane incompletely inhibits fluid flow across the three or more microfluidic channels.
  5. The microfluidic device of claim 1 wherein the diaphragm valve further comprises a vias layer.
  6. The microfluidic device of claim 1, wherein applying pressure or vacuum to the one or more drive channels regulates the flow of fluid across the elastomeric membrane to form three or more channel valves.
  7. The microfluidic device of claim 1 wherein the membrane naturally closes the valve and applying a vacuum to the membrane deflects the membrane from the valve seat and opens the valve.
  8. The microfluidic device of claim 1, wherein the microfluidic layer comprises a surface facing the membrane, the surface comprising a groove forming a microfluidic channel when pressed against the membrane.
  9. The microfluidic device of claim 1 wherein the microfluidic layer comprises internal microfluidic channels that intersect in holes in the layer that are open to a nexus.
  10. The microfluidic device of claim 1, wherein the drive layer comprises a surface against the membrane, the surface comprising a groove forming a drive channel when pressed against the membrane.
  11. The microfluidic device of claim 1 wherein the drive layer includes an internal drive channel that opens to the valve chamber.
  12. 7. The microfluidic device of claim 6 wherein the three or more microfluidic channels gather in a Y-shaped nexus.
  13. The microfluidic device of claim 1 comprising a plurality of said diaphragm valves driven by a single drive channel.
  14. A microfluidic device comprising a microfluidic layer, a drive layer and an elastomeric layer sandwiched therebetween,
    The microfluidic layer comprises first and second microfluidic channels gathered at a nexus and separated by discontinuities that impede fluid flow between the first and second channels but not along the second channel,
    The drive layer includes one or more drive channels that open into a valve chamber disposed opposite the nexus,
    And a diaphragm valve is formed by adjusting the flow of fluid across the three or more microfluidic channels by displacement of the elastomeric membrane.
  15. 15. The microfluidic device of claim 14 wherein the fluid flow in the second microfluidic channel is not regulated by the elastomeric membrane.
  16. 15. The microfluidic device of claim 14 wherein the elastomeric membrane disrupts fluid flow across one or more microfluidic channels.
  17. 15. The microfluidic device of claim 14 wherein the elastomeric membrane incompletely inhibits fluid flow across one or more microfluidic channels.
  18. 15. The microfluidic device of claim 14 wherein the diaphragm valve further comprises a valve layer.
  19. 19. The microfluidic device of claim 18 wherein applying pressure or vacuum to the one or more drive channels regulates the fluid flow across the one or more discontinuities.
  20. 19. The microfluidic device of claim 18 wherein the two or more microfluidic channels gather in a T-shaped nexus.
  21. 15. The microfluidic device of claim 14 comprising a plurality of said diaphragm valves driven by a single drive channel.
  22. 15. The microfluidic device of claim 14, wherein the second channel forms a loop having a predetermined volume and comprises a predetermined volume of positive displacement pump.
  23. A method comprising pumping a predetermined volume of liquid into a first channel of the apparatus of claim 22 by opening the diaphragm valve and performing a plurality of strokes using a pump.
  24. A microfluidic device comprising a microfluidic layer, a drive layer and an elastomeric layer sandwiched therebetween,
    The microfluidic layer comprises a microfluidic channel,
    The drive layer comprises one or more drive channels that open to a valve chamber disposed along the microfluidic channel,
    The fluid flows along a channel whether the elastomeric membrane is displaced or not, but the displacement of the elastomeric membrane regulates fluid flow along the channel to form a diaphragm valve.
  25. 25. The microfluidic device of claim 24 wherein the elastomeric membrane modulates fluid flow by increasing or decreasing the cross-sectional area of the one or more microchannels.
  26. 25. The microfluidic device of claim 24 wherein the diaphragm valve further comprises a bias layer.
  27. 25. The microfluidic device of claim 24 wherein applying pressure or vacuum to the one or more drive channels regulates the flow of fluid across the one or more discontinuities.
  28. 25. The microfluidic device of claim 24 further comprising a magnet for fielding a diaphragm valve.
  29. 29. The magnet of claim 28, wherein the magnet is selected from the group consisting of permanent magnets, electromagnets, and rare earth magnets.
  30. 25. The microfluidic device of claim 24 wherein the path of the one or more microchannels forms a star or nested star adjacent the diaphragm valve.
  31. A system comprising a first microfluidic device and a second microfluidic device,
    The first microfluidic device comprises an inlet, an outlet, a pump configured to pump fluid through the circuit, and a reactor, capture zone, temperature circulation zone, hot zone, cold zone, separation channel, analysis circuit, mixer, bead processing A first microfluidic circuit comprising a bead processing unit and at least one first functional component selected from a magnet,
    The second microfluidic device comprises a plurality of second microfluidic circuits each comprising an inlet, an outlet, and one or more functional components different from the first functional component,
    The first component and the second component are configured to occupy a plurality of positions, in each of which the outlet of the first circuit is paired with an inlet of one of the second circuits from the first circuit to the paired second circuit. System that allows fluids to flow.
  32. 32. The system of claim 31, wherein the first microfluidic device further comprises an electrode configured to electrically move the fluid, molecule, chemical or particle.
  33. 32. The system of claim 31, wherein said second microfluidic device further comprises an analysis region.
  34. 32. The system of claim 31, wherein said second microfluidic device further comprises a capture region.
  35. 32. The system of claim 31, wherein said second microfluidic device is mobile relative to said first microfluidic device.
  36. 32. The system of claim 31, wherein the fluid input of the second microfluidic device transfers fluid from the first microfluidic device to a plurality of microfluidic channels.
  37. 32. The system of claim 31, wherein the fluid output of the first microfluidic device delivers fluid from a plurality of microfluidic channels to the fluid input of the second microfluidic device.
  38. 33. The system of claim 32, wherein the fluid, molecule, chemical or particle is moved by electrophoresis from the fluid output of the first microfluidic device to the fluid input of the second microfluidic device.
  39. a) performing a first operation on a first sample in a first microfluidic circuit of the first microfluidic device of claim 26;
    b) engaging the first and second microfluidic devices of claim 31 such that the output of the first circuit is mated with the inlet of the first portion of the second microfluidic circuit;
    c) moving the first sample from the first circuit to the first portion of the second circuit after operation;
    d) performing a second operation on a first sample received in a first portion of a second circuit;
    e) performing a first operation on a second sample in a first microfluidic circuit;
    f) engaging the first and second microfluidic devices such that the output of the first circuit is paired with the inlet of the next circuit that is different from the second microfluidic circuit;
    g) moving the second sample from the first circuit to the next of the second circuit after operation; And
    h) performing a second operation on the first reaction sample in the first portion of the second circuit
    How to include.
  40. 40. The method of claim 39, comprising repeating steps e) to f) for one or more samples in the first circuit, wherein each sample is moved to a next circuit different from the plurality of second circuits. .
  41. The method of claim 39, wherein the first operation comprises mixing the sample with a reagent.
  42. 40. The method of claim 39, wherein the first operation is performed in less time than the second operation.
  43. A method of making a microfluidic device, comprising connecting multiple layers to form a plurality of microchannels and diaphragm valves, the method comprising:
    The multiple layers are sandwiched together;
    At least two of the plurality of layers are selected from the group consisting of elastomeric membranes, drive layers, microfluidic layers, valve layers, heat spreaders, bias layers, interface layers, and cover layers.
  44. The method of claim 43, wherein at least one of the plurality of layers is an adhesive layer.
  45. 44. The method of claim 43, wherein the multiple layers are connected together by an adhesive layer.
  46. The method of claim 43, wherein the multiple layers are connected together by one or more clamps.
  47. The method of claim 43, wherein the multiple layers are connected except for the diaphragm valve position by reducing pressure or temperature at the valve.
  48. The method of claim 43, wherein the multiple layers are connected except for the diaphragm valve position by selectively placing a coating on the valve.
  49. The method of claim 48, wherein the coating is removable.
  50. a) microfluidic channel;
    b) a first temperature zone disposed along a channel having a temperature above ambient temperature;
    c) a second temperature zone disposed along the channel having a temperature below ambient temperature; And
    d) a volumetric pump disposed along the channel and configured to pump the liquid into the first and second temperature zones
    Microfluidic device comprising a.
  51. 51. The microfluidic device of claim 50, further comprising a microfluidic loop thermally coupled to at least one temperature zone.
  52. 51. The microfluidic device of claim 50, wherein the samples in the one or more microfluidic channels are pumped two or more times between two or more different temperature zones.
  53. 51. The microfluidic device of claim 50 wherein said sample is separated from at least one three valve pump by an immiscible fluid.
  54. 51. The microfluidic device of claim 50 wherein the at least one diaphragm valve is located in a temperature zone.
  55. Heating or cooling the nucleic acid sequence in the microfluidic device of claim 41; And
    Analyzing the nucleic acid sequence
    Including, the analysis method of the sample.
  56. The method of claim 55, wherein said analysis comprises sequencing, ligation or polymerase chain reaction amplification, transcription, translation, or co-transcriptional translation.
  57. The method of claim 55, wherein the nucleic acid is selected from the group consisting of genomic DNA, mitochondrial DNA, mRNA, tRNA, rRNA, siRNA.
  58. The method of claim 55, wherein said analysis comprises ligation of at least one adapter to said nucleic acid sequence.
  59. The method of claim 55, wherein the adapter comprises a specific nucleic acid sequence identifier.
  60. The method of claim 55, wherein the specific nucleic acid sequence identifier is used as an internal quality control metric.
  61. The method of claim 55, wherein said analysis comprises binding of a single nucleic acid sequence ligated to a bead.
  62. The method of claim 61, further comprising amplifying said nucleic acid sequence.
  63. A microfluidic layer comprising microfluidic channels; And
    Magnetic field generating means for generating a magnetic field in the tight bend region
    As a microfluidic device comprising:
    The channel comprises at least one tight bend comprising two channel segments connected to each other and oriented at an acute angle,
    Paramagnetic particles flowing through the close bend is retarded by a magnetic field.
  64. 64. The microfluidic device of claim 63 comprising a plurality of tight bends.
  65. A microfluidic layer comprising microfluidic channels; And
    Magnetic field generating means for generating a magnetic field in the second region
    As a microfluidic device comprising:
    The channel in turn comprises first, second and third regions, the second region having a larger cross-sectional area than the first and third regions,
    A microfluidic device in which paramagnetic particles flowing through a second region are retarded by a magnetic field.
  66. Mixing the biomolecule and reagent in a first reaction chamber on the microfluidic device to produce a first reaction mixture;
    Moving the reaction mixture to a reaction zone in the apparatus and carrying out the reaction to produce a product mixture;
    Moving the product mixture to a region in the device to capture the product on paramagnetic capture particles within the region;
    Moving the particles and the trapped product to a capture chamber in the device in a magnetic field to receive the trapped particles and product in the capture chamber;
    Cleaning the particles in the capture chamber; And
    Moving the particles and product to a port on the device, wherein the product can be withdrawn from the device
    Including, the method of performing biomolecular analysis on the microfluidic device.
  67. The method of claim 66, wherein the biomolecule is selected from nucleic acids, proteins, carbohydrates, cells, or lipids.
  68. 67. The method of claim 66, wherein the reaction is a nucleic acid amplification reaction.
  69. The method of claim 68, wherein the amplification is isothermal.
  70. 69. The method of claim 68, wherein the amplification comprises thermal cycling of the mixture.
  71. 67. The method of claim 66, further comprising performing a second reaction on the reaction mixture or product mixture.
  72. a) each
    i) two or more ports configured to receive a sample or to remove a sample therefrom;
    ii) at least one pump configured to pump fluid through the circuit;
    iii) one or more reaction chambers comprising means for conducting a chemical or biochemical reaction; And
    iv) at least one capture chamber comprising means for capturing particles
    A plurality of microfluidic reaction circuit comprising a; And
    b) one or more dispensing ports in fluid communication with a plurality of microfluidic circuits and configured to deliver sample or reagent fluid to each of the circuits;
    As a microfluidic device comprising:
    And the plurality of circuits are configured to operate simultaneously on a plurality of different materials delivered to one of the ports in each circuit.
  73. 73. The method of claim 72, wherein the reaction chamber is configured to be heated or cooled by a heat pump.
  74. The method of claim 72, wherein the capture chamber is disposed in a magnetic field configured to retard the movement of paramagnetic particles in the capture chamber.
  75. 73. The method of claim 72, wherein the pump is a volumetric pump.
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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011133014A1 (en) * 2010-04-19 2011-10-27 Mimos Berhad Planar micropump with integrated microvalves
WO2012144794A2 (en) * 2011-04-19 2012-10-26 주식회사 바이오포커스 Method for manufacturing a microvalve device mounted on a lab-on-a-chip, and microvalve device manufactured by same
KR101299438B1 (en) * 2011-04-19 2013-08-29 손문탁 method for manufacturing microvalve element mounted on lab-on-a-chip, and microvalve element produced thereby
KR101404657B1 (en) * 2013-04-26 2014-06-09 주식회사 바이오록스 Manifold Package for Driving Lab-on-a-Chip That Can Supply Vacuum, Air Vent and Compressed Air
US8778282B2 (en) 2010-06-15 2014-07-15 Samsung Electronics Co., Ltd. Microfluidic device having microvalve

Families Citing this family (96)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6432290B1 (en) 1999-11-26 2002-08-13 The Governors Of The University Of Alberta Apparatus and method for trapping bead based reagents within microfluidic analysis systems
EP1594694A4 (en) * 2002-12-30 2010-01-20 Univ California Methods and apparatus for pathogen detection and analysis
US7799553B2 (en) * 2004-06-01 2010-09-21 The Regents Of The University Of California Microfabricated integrated DNA analysis system
WO2006032044A2 (en) 2004-09-15 2006-03-23 Microchip Biotechnologies, Inc. Microfluidic devices
GB0421529D0 (en) 2004-09-28 2004-10-27 Landegren Gene Technology Ab Microfluidic structure
EP1984738A2 (en) 2006-01-11 2008-10-29 Raindance Technologies, Inc. Microfluidic devices and methods of use in the formation and control of nanoreactors
KR20080096567A (en) 2006-02-03 2008-10-30 마이크로칩 바이오테크놀로지스, 인크. Microfluidic devices
US7766033B2 (en) 2006-03-22 2010-08-03 The Regents Of The University Of California Multiplexed latching valves for microfluidic devices and processors
US9562837B2 (en) 2006-05-11 2017-02-07 Raindance Technologies, Inc. Systems for handling microfludic droplets
WO2008052138A2 (en) 2006-10-25 2008-05-02 The Regents Of The University Of California Inline-injection microdevice and microfabricated integrated dna analysis system using same
EP2109666A4 (en) 2007-02-05 2011-09-14 Integenx Inc Microfluidic and nanofluidic devices, systems, and applications
WO2008097559A2 (en) 2007-02-06 2008-08-14 Brandeis University Manipulation of fluids and reactions in microfluidic systems
US8592221B2 (en) 2007-04-19 2013-11-26 Brandeis University Manipulation of fluids, fluid components and reactions in microfluidic systems
US8454906B2 (en) 2007-07-24 2013-06-04 The Regents Of The University Of California Microfabricated droplet generator for single molecule/cell genetic analysis in engineered monodispersed emulsions
WO2009078016A2 (en) 2007-12-17 2009-06-25 Yeda Research And Develompment Co. Ltd. System and method for editing and manipulating dna
CN101990516B (en) 2008-01-22 2015-09-09 英特基因有限公司 Multiplex sample preparation system and the use in integrated analysis system thereof
US20120171680A1 (en) * 2008-06-12 2012-07-05 Shapiro Ehud Y Single-molecule pcr for amplification from a single nucleotide strand
WO2010023596A1 (en) * 2008-08-25 2010-03-04 Koninklijke Philips Electronics N.V. Reconfigurable microfluidic filter
WO2010077322A1 (en) 2008-12-31 2010-07-08 Microchip Biotechnologies, Inc. Instrument with microfluidic chip
NZ620826A (en) 2009-02-03 2015-09-25 Netbio Inc Nucleic acid purification
EP2414845A2 (en) * 2009-04-02 2012-02-08 Purdue Research Foundation, Inc. Variable volume mixing and automatic fluid management for programmable microfluidics
US9357946B2 (en) * 2009-04-15 2016-06-07 Nanomix, Inc. Breath condensate sampler and detector and breath/breath condensate sampler and detector
WO2010141326A1 (en) 2009-06-02 2010-12-09 Integenx Inc. Fluidic devices with diaphragm valves
US20110065101A1 (en) 2009-06-04 2011-03-17 Lockheed Martin Corporation Multiple-sample microfluidic chip for DNA analysis
SG176669A1 (en) 2009-06-05 2012-01-30 Integenx Inc Universal sample preparation system and use in an integrated analysis system
US9500645B2 (en) 2009-11-23 2016-11-22 Cyvek, Inc. Micro-tube particles for microfluidic assays and methods of manufacture
US9700889B2 (en) 2009-11-23 2017-07-11 Cyvek, Inc. Methods and systems for manufacture of microarray assay systems, conducting microfluidic assays, and monitoring and scanning to obtain microfluidic assay results
US9855735B2 (en) 2009-11-23 2018-01-02 Cyvek, Inc. Portable microfluidic assay devices and methods of manufacture and use
WO2013134740A1 (en) 2012-03-08 2013-09-12 Cyvek, Inc. Methods and systems for epi-fluorescent monitoring and scanning for microfluidic assays
CA2781556C (en) 2009-11-23 2015-05-19 Cyvek, Inc. Method and apparatus for performing assays
US10022696B2 (en) 2009-11-23 2018-07-17 Cyvek, Inc. Microfluidic assay systems employing micro-particles and methods of manufacture
US9759718B2 (en) 2009-11-23 2017-09-12 Cyvek, Inc. PDMS membrane-confined nucleic acid and antibody/antigen-functionalized microlength tube capture elements, and systems employing them, and methods of their use
US10065403B2 (en) 2009-11-23 2018-09-04 Cyvek, Inc. Microfluidic assay assemblies and methods of manufacture
US8584703B2 (en) 2009-12-01 2013-11-19 Integenx Inc. Device with diaphragm valve
IT1397819B1 (en) 2009-12-17 2013-02-04 Silicon Biosystems Spa microfluidic system
IT1397820B1 (en) 2009-12-17 2013-02-04 Silicon Biosystems Spa microfluidic system
IT1398480B1 (en) 2009-12-17 2013-03-01 Silicon Biosystems Spa microfluidic system
US9074242B2 (en) 2010-02-12 2015-07-07 Raindance Technologies, Inc. Digital analyte analysis
US9366632B2 (en) 2010-02-12 2016-06-14 Raindance Technologies, Inc. Digital analyte analysis
US10351905B2 (en) 2010-02-12 2019-07-16 Bio-Rad Laboratories, Inc. Digital analyte analysis
ITTO20100319A1 (en) * 2010-04-20 2011-10-21 Eltek Spa microfluidic devices and / or equipment for microfluidic devices
ITTO20100068U1 (en) 2010-04-20 2011-10-21 Eltek Spa microfluidic devices and / or equipment for microfluidic devices
US8512538B2 (en) 2010-05-28 2013-08-20 Integenx Inc. Capillary electrophoresis device
GB2481425A (en) 2010-06-23 2011-12-28 Iti Scotland Ltd Method and device for assembling polynucleic acid sequences
KR20120015593A (en) * 2010-08-12 2012-02-22 삼성전자주식회사 Microfluidic device having microvalve
WO2012024658A2 (en) 2010-08-20 2012-02-23 IntegenX, Inc. Integrated analysis system
EP2606242A4 (en) 2010-08-20 2016-07-20 Integenx Inc Microfluidic devices with mechanically-sealed diaphragm valves
US8961764B2 (en) 2010-10-15 2015-02-24 Lockheed Martin Corporation Micro fluidic optic design
TWI532530B (en) * 2010-10-29 2016-05-11 萬國商業機器公司 Multilayer microfluidic probe head with immersion channels and fabrication thereof
CA2830533A1 (en) 2011-03-22 2012-09-27 Cyvek, Inc. Microfluidic devices and methods of manufacture and use
US8841071B2 (en) * 2011-06-02 2014-09-23 Raindance Technologies, Inc. Sample multiplexing
CN104040319A (en) * 2011-06-14 2014-09-10 康宁股份有限公司 Hybrid microfluidic assemblies
DE102011078770B4 (en) * 2011-07-07 2016-04-28 Robert Bosch Gmbh Microfluidic device, microfluidic system and method of transporting fluids
JP6068850B2 (en) 2011-07-25 2017-01-25 株式会社エンプラス Fluid handling apparatus and fluid handling method
US20150136604A1 (en) 2011-10-21 2015-05-21 Integenx Inc. Sample preparation, processing and analysis systems
US8894946B2 (en) 2011-10-21 2014-11-25 Integenx Inc. Sample preparation, processing and analysis systems
US10222391B2 (en) 2011-12-07 2019-03-05 The Johns Hopkins University System and method for screening a library of samples
US9101930B2 (en) 2012-02-13 2015-08-11 Neumodx Molecular, Inc. Microfluidic cartridge for processing and detecting nucleic acids
US9637775B2 (en) 2012-02-13 2017-05-02 Neumodx Molecular, Inc. System and method for processing biological samples
US9604213B2 (en) 2012-02-13 2017-03-28 Neumodx Molecular, Inc. System and method for processing and detecting nucleic acids
US9322054B2 (en) 2012-02-22 2016-04-26 Lockheed Martin Corporation Microfluidic cartridge
US9165097B2 (en) * 2012-03-08 2015-10-20 Purdue Research Foundation Programmable microfluidic systems and related methods
US9528082B2 (en) 2012-07-25 2016-12-27 The Charles Stark Draper Laboratory, Inc. Modular platform for multi-tissue integrated cell culture
DE102012217487A1 (en) * 2012-09-26 2014-04-17 Agilent Technologies, Inc. - A Delaware Corporation - Fluid interface between fluid lines of different cross sections
US9192934B2 (en) * 2012-10-25 2015-11-24 General Electric Company Insert assembly for a microfluidic device
US10215687B2 (en) 2012-11-19 2019-02-26 The General Hospital Corporation Method and system for integrated mutliplexed photometry module
EP2920574A4 (en) * 2012-11-19 2016-09-28 Gen Hospital Corp System and method for integrated multiplexed photometry module
WO2014093360A1 (en) * 2012-12-10 2014-06-19 Landers James P Frequency-based filtering of mechanical actuation using fluidic device
US9249387B2 (en) * 2013-01-29 2016-02-02 The Charles Stark Draper Laboratory, Inc. Modular platform for multi-tissue integrated cell culture
EP2848698A1 (en) * 2013-08-26 2015-03-18 F. Hoffmann-La Roche AG System and method for automated nucleic acid amplification
EP3046663A4 (en) * 2013-09-18 2017-05-31 California Institute of Technology System and method for movement and timing control
CN110560187A (en) 2013-11-18 2019-12-13 尹特根埃克斯有限公司 Cartridge and instrument for sample analysis
US20150166956A1 (en) * 2013-12-16 2015-06-18 General Electric Company Devices for separation of particulates, associated methods and systems
US10195610B2 (en) 2014-03-10 2019-02-05 Click Diagnostics, Inc. Cartridge-based thermocycler
WO2015179098A1 (en) 2014-05-21 2015-11-26 Integenx Inc. Fluidic cartridge with valve mechanism
US10210410B2 (en) 2014-10-22 2019-02-19 Integenx Inc. Systems and methods for biometric data collections
CN104568287B (en) * 2014-12-24 2017-07-14 北京工业大学 Pressure apparatus in a kind of deformation direct measurement microchannel of utilization PDMS film
EP3240906A4 (en) 2014-12-31 2018-09-05 Click Diagnostics, Inc. Devices and methods for molecular diagnostic testing
US10048190B2 (en) * 2015-01-21 2018-08-14 Sbt Instruments Aps Microfluidic particle analysis device
CN107454862A (en) 2015-02-04 2017-12-08 查尔斯斯塔克布料实验室公司 The actuating valve or pump of microfluidic device
US9868659B2 (en) 2015-04-17 2018-01-16 General Electric Company Subsurface water purification method
US10233491B2 (en) 2015-06-19 2019-03-19 IntegenX, Inc. Valved cartridge and system
US10519493B2 (en) 2015-06-22 2019-12-31 Fluxergy, Llc Apparatus and method for image analysis of a fluid sample undergoing a polymerase chain reaction (PCR)
US10214772B2 (en) 2015-06-22 2019-02-26 Fluxergy, Llc Test card for assay and method of manufacturing same
JP2018528845A (en) 2015-09-14 2018-10-04 メディシーブ リミテッド Magnetic filtration apparatus and method
US10228367B2 (en) 2015-12-01 2019-03-12 ProteinSimple Segmented multi-use automated assay cartridge
CN105567548B (en) * 2015-12-17 2018-08-10 青岛意诚融智生物仪器有限公司 A kind of micro-fluidic chip and detection method for quick multiplexed PCR amplification
EP3338062B1 (en) * 2016-01-22 2019-11-20 Hewlett-Packard Development Company, L.P. Fluid sensing with control of particle aggregation in sensing zone
US20190176122A1 (en) * 2016-05-02 2019-06-13 Purdue Research Foundation Systems and methods for producing a chemical product
EP3523048A4 (en) * 2016-10-06 2019-08-14 Univ California Volumetric micro-injector for capillary electrophoresis
DE102016222032A1 (en) * 2016-11-10 2018-05-17 Robert Bosch Gmbh Microfluidic device and method for analyzing nucleic acids
CN108217576A (en) * 2016-12-21 2018-06-29 上海新微技术研发中心有限公司 Diaphragm stop valve and its manufacturing method
WO2019089268A1 (en) * 2017-10-23 2019-05-09 Conservation X Labs, Inc. Systems and methods relating to portable microfluidic devices for processing biomolecules
DE102018204633A1 (en) * 2018-03-27 2019-10-02 Robert Bosch Gmbh Microfluidic device and method for processing a fluid
WO2019204279A1 (en) * 2018-04-16 2019-10-24 Klaris Corporation Methods and apparatus for forming 2-dimensional drop arrays
WO2019226682A1 (en) * 2018-05-22 2019-11-28 California Institute Of Technology Miniature fixed and adjustable flow restrictor for the body

Family Cites Families (335)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3190310A (en) 1960-12-16 1965-06-22 American Radiator & Standard Gang valve arrangement
US3352643A (en) 1964-09-02 1967-11-14 Hitachi Ltd Liquid chromatography and chromatographs
US3433257A (en) * 1966-02-01 1969-03-18 Ibm Diaphragm type fluid logic latch
US3568692A (en) * 1967-11-27 1971-03-09 Bowles Eng Corp Optical machining process
US3610274A (en) 1969-10-08 1971-10-05 Brown & Sharpe Mfg Fluid logic circuit
US4113665A (en) 1977-02-03 1978-09-12 Ameron, Inc. Coatings prepared from trialkoxysilanes
US4558845A (en) 1982-09-22 1985-12-17 Hunkapiller Michael W Zero dead volume valve
US4703913A (en) 1982-09-22 1987-11-03 California Institute Of Technology Diaphragm valve
US4963498A (en) 1985-08-05 1990-10-16 Biotrack Capillary flow device
GB8523166D0 (en) 1985-09-19 1985-10-23 Yarsley Technical Centre Ltd Scratch resistant coatings
US5085757A (en) * 1987-11-25 1992-02-04 Northeastern University Integrated temperature control/alignment system for high performance capillary electrophoretic apparatus
US5523231A (en) 1990-02-13 1996-06-04 Amersham International Plc Method to isolate macromolecules using magnetically attractable beads which do not specifically bind the macromolecules
US5750015A (en) 1990-02-28 1998-05-12 Soane Biosciences Method and device for moving molecules by the application of a plurality of electrical fields
US6176962B1 (en) * 1990-02-28 2001-01-23 Aclara Biosciences, Inc. Methods for fabricating enclosed microchannel structures
US5387505A (en) * 1990-05-04 1995-02-07 Eastman Kodak Company Preparation and isolation of single-stranded biotinylated nucleic acids by heat avidin-biotin cleavage
SE470347B (en) 1990-05-10 1994-01-31 Pharmacia Lkb Biotech Microstructure fluid flow system and method for manufacturing such systems
DE69104775D1 (en) 1990-05-29 1994-12-01 Waters Investments Ltd Method and apparatus for performing capillary electrophoresis.
US5364759B2 (en) 1991-01-31 1999-07-20 Baylor College Medicine Dna typing with short tandem repeat polymorphisms and identification of polymorphic short tandem repeats
US5726026A (en) * 1992-05-01 1998-03-10 Trustees Of The University Of Pennsylvania Mesoscale sample preparation device and systems for determination and processing of analytes
US5587128A (en) 1992-05-01 1996-12-24 The Trustees Of The University Of Pennsylvania Mesoscale polynucleotide amplification devices
AT140880T (en) 1992-05-01 1996-08-15 Univ Pennsylvania Fluessigkeitsbehandlung in microfabricated analytical devices
DE69533159T2 (en) 1994-11-14 2005-07-21 Trustees Of The University Of Pennsylvania Miniaturized sample preparation devices and systems for determining and treating analytes
US5304487A (en) 1992-05-01 1994-04-19 Trustees Of The University Of Pennsylvania Fluid handling in mesoscale analytical devices
US5637469A (en) 1992-05-01 1997-06-10 Trustees Of The University Of Pennsylvania Methods and apparatus for the detection of an analyte utilizing mesoscale flow systems
US5275645A (en) * 1992-11-24 1994-01-04 Ameron, Inc. Polysiloxane coating
US5482836A (en) * 1993-01-14 1996-01-09 The Regents Of The University Of California DNA purification by triplex-affinity capture and affinity capture electrophoresis
DE69333601T2 (en) 1993-04-15 2005-09-15 Zeptosens Ag Method of controlling sample introduction in microseparation techniques and sampling devices
US5482845A (en) 1993-09-24 1996-01-09 The Trustees Of Columbia University In The City Of New York Method for construction of normalized cDNA libraries
US5776748A (en) 1993-10-04 1998-07-07 President And Fellows Of Harvard College Method of formation of microstamped patterns on plates for adhesion of cells and other biological materials, devices and uses therefor
US5453163A (en) 1993-10-29 1995-09-26 Yan; Chao Electrokinetic packing of capillary columns
US6358385B1 (en) * 1993-12-17 2002-03-19 The Perkin-Elmer Corporation Polymers for separation of biomolecules by capillary electrophoresis
US5639428A (en) 1994-07-19 1997-06-17 Becton Dickinson And Company Method and apparatus for fully automated nucleic acid amplification, nucleic acid assay and immunoassay
US6001229A (en) * 1994-08-01 1999-12-14 Lockheed Martin Energy Systems, Inc. Apparatus and method for performing microfluidic manipulations for chemical analysis
US5705628A (en) * 1994-09-20 1998-01-06 Whitehead Institute For Biomedical Research DNA purification and isolation using magnetic particles
US5571410A (en) 1994-10-19 1996-11-05 Hewlett Packard Company Fully integrated miniaturized planar liquid sample handling and analysis device
US5775371A (en) 1995-03-08 1998-07-07 Abbott Laboratories Valve control
US5741462A (en) * 1995-04-25 1998-04-21 Irori Remotely programmable matrices with memories
US5589136A (en) * 1995-06-20 1996-12-31 Regents Of The University Of California Silicon-based sleeve devices for chemical reactions
US6168948B1 (en) * 1995-06-29 2001-01-02 Affymetrix, Inc. Miniaturized genetic analysis systems and methods
US5856174A (en) * 1995-06-29 1999-01-05 Affymetrix, Inc. Integrated nucleic acid diagnostic device
US5872010A (en) 1995-07-21 1999-02-16 Northeastern University Microscale fluid handling system
US20020068357A1 (en) 1995-09-28 2002-06-06 Mathies Richard A. Miniaturized integrated nucleic acid processing and analysis device and method
US5705813A (en) * 1995-11-01 1998-01-06 Hewlett-Packard Company Integrated planar liquid handling system for maldi-TOF MS
EP0779512B1 (en) 1995-12-14 2001-10-31 Hewlett-Packard Company, A Delaware Corporation Column for capillary chromatographic separations
US5863502A (en) * 1996-01-24 1999-01-26 Sarnoff Corporation Parallel reaction cassette and associated devices
US5948684A (en) 1997-03-31 1999-09-07 University Of Washington Simultaneous analyte determination and reference balancing in reference T-sensor devices
US5994064A (en) 1996-04-24 1999-11-30 Identigene, Inc. Simple and complex tandem repeats with DNA typing method
US6387707B1 (en) 1996-04-25 2002-05-14 Bioarray Solutions Array Cytometry
AU3570797A (en) 1996-06-14 1998-01-07 University Of Washington Absorption-enhanced differential extraction device
US5942443A (en) 1996-06-28 1999-08-24 Caliper Technologies Corporation High throughput screening assay systems in microscale fluidic devices
US6267858B1 (en) 1996-06-28 2001-07-31 Caliper Technologies Corp. High throughput screening assay systems in microscale fluidic devices
US6074827A (en) 1996-07-30 2000-06-13 Aclara Biosciences, Inc. Microfluidic method for nucleic acid purification and processing
US5770029A (en) 1996-07-30 1998-06-23 Soane Biosciences Integrated electrophoretic microdevices
US6136212A (en) 1996-08-12 2000-10-24 The Regents Of The University Of Michigan Polymer-based micromachining for microfluidic devices
US6129828A (en) 1996-09-06 2000-10-10 Nanogen, Inc. Apparatus and methods for active biological sample preparation
US5935401A (en) 1996-09-18 1999-08-10 Aclara Biosciences Surface modified electrophoretic chambers
US6110343A (en) 1996-10-04 2000-08-29 Lockheed Martin Energy Research Corporation Material transport method and apparatus
AU746549B2 (en) * 1996-11-20 2002-05-02 Becton Dickinson & Company Microfabricated isothermal nucleic acid amplification devices and methods
US6306588B1 (en) * 1997-02-07 2001-10-23 Invitrogen Corporation Polymerases for analyzing or typing polymorphic nucleic acid fragments and uses thereof
US6117634A (en) * 1997-03-05 2000-09-12 The Reagents Of The University Of Michigan Nucleic acid sequencing and mapping
WO1998045684A1 (en) * 1997-04-04 1998-10-15 Biosite Diagnostics, Inc. Methods for concentrating ligands using magnetic particles
US6235471B1 (en) 1997-04-04 2001-05-22 Caliper Technologies Corp. Closed-loop biochemical analyzers
US5885470A (en) * 1997-04-14 1999-03-23 Caliper Technologies Corporation Controlled fluid transport in microfabricated polymeric substrates
US6872527B2 (en) 1997-04-16 2005-03-29 Xtrana, Inc. Nucleic acid archiving
US6632619B1 (en) 1997-05-16 2003-10-14 The Governors Of The University Of Alberta Microfluidic system and methods of use
AU734957B2 (en) 1997-05-16 2001-06-28 Alberta Research Council Inc. Microfluidic system and methods of use
WO1998053300A2 (en) 1997-05-23 1998-11-26 Lynx Therapeutics, Inc. System and apparaus for sequential processing of analytes
US5900130A (en) 1997-06-18 1999-05-04 Alcara Biosciences, Inc. Method for sample injection in microchannel device
GB9713597D0 (en) 1997-06-28 1997-09-03 Sec Dep Of The Home Department Improvements in and relating to forensic identification
US6073482A (en) 1997-07-21 2000-06-13 Ysi Incorporated Fluid flow module
US6190616B1 (en) * 1997-09-11 2001-02-20 Molecular Dynamics, Inc. Capillary valve, connector, and router
US6207031B1 (en) * 1997-09-15 2001-03-27 Whitehead Institute For Biomedical Research Methods and apparatus for processing a sample of biomolecular analyte using a microfabricated device
US6007775A (en) 1997-09-26 1999-12-28 University Of Washington Multiple analyte diffusion based chemical sensor
US5842787A (en) 1997-10-09 1998-12-01 Caliper Technologies Corporation Microfluidic systems incorporating varied channel dimensions
US6803019B1 (en) 1997-10-15 2004-10-12 Aclara Biosciences, Inc. Laminate microstructure device and method for making same
US6120985A (en) 1997-10-31 2000-09-19 Bbi Bioseq, Inc. Pressure-enhanced extraction and purification
US5860594A (en) * 1997-12-19 1999-01-19 Carrier Corporation Method and apparatus for changing operational modes of a transport refrigeration system
US6200814B1 (en) 1998-01-20 2001-03-13 Biacore Ab Method and device for laminar flow on a sensing surface
US6167910B1 (en) 1998-01-20 2001-01-02 Caliper Technologies Corp. Multi-layer microfluidic devices
US6251343B1 (en) 1998-02-24 2001-06-26 Caliper Technologies Corp. Microfluidic devices and systems incorporating cover layers
WO1999046591A2 (en) 1998-03-10 1999-09-16 Strategic Diagnostics, Inc. Integrated assay device and methods of production and use
US6979424B2 (en) * 1998-03-17 2005-12-27 Cepheid Integrated sample analysis device
WO1999058664A1 (en) * 1998-05-14 1999-11-18 Whitehead Institute For Biomedical Research Solid phase technique for selectively isolating nucleic acids
US6787111B2 (en) 1998-07-02 2004-09-07 Amersham Biosciences (Sv) Corp. Apparatus and method for filling and cleaning channels and inlet ports in microchips used for biological analysis
US6627446B1 (en) 1998-07-02 2003-09-30 Amersham Biosciences (Sv) Corp Robotic microchannel bioanalytical instrument
US6787308B2 (en) 1998-07-30 2004-09-07 Solexa Ltd. Arrayed biomolecules and their use in sequencing
US6387234B1 (en) 1998-08-31 2002-05-14 Iowa State University Research Foundation, Inc. Integrated multiplexed capillary electrophoresis system
US6103199A (en) 1998-09-15 2000-08-15 Aclara Biosciences, Inc. Capillary electroflow apparatus and method
US6572830B1 (en) 1998-10-09 2003-06-03 Motorola, Inc. Integrated multilayered microfludic devices and methods for making the same
GB9900298D0 (en) 1999-01-07 1999-02-24 Medical Res Council Optical sorting method
US6685809B1 (en) * 1999-02-04 2004-02-03 Ut-Battelle, Llc Methods for forming small-volume electrical contacts and material manipulations with fluidic microchannels
CA2359901A1 (en) * 1999-02-16 2000-08-24 Pe Corporation (Ny) Bead dispensing system
WO2000050172A1 (en) 1999-02-23 2000-08-31 Caliper Technologies Corp. Manipulation of microparticles in microfluidic systems
AU772213C (en) 1999-02-26 2004-12-09 Exact Sciences Corporation Biochemical purification devices with immobilized capture probes and their uses
EP1411340A3 (en) 1999-02-26 2004-05-19 EXACT Sciences Corporation Biochemical purification devices with immobilized capture probes and their uses
US6319476B1 (en) 1999-03-02 2001-11-20 Perseptive Biosystems, Inc. Microfluidic connector
US7150994B2 (en) * 1999-03-03 2006-12-19 Symyx Technologies, Inc. Parallel flow process optimization reactor
US6048100A (en) * 1999-03-10 2000-04-11 Industrial Label Corp. Resealable closure for a bag
US6994986B2 (en) * 1999-03-17 2006-02-07 The Board Of Trustees Of The Leland Stanford University In vitro synthesis of polypeptides by optimizing amino acid metabolism
AU4226900A (en) 1999-04-08 2000-10-23 Joseph L. Chan Apparatus for fast preparation and analysis of nucleic acids
US6322683B1 (en) 1999-04-14 2001-11-27 Caliper Technologies Corp. Alignment of multicomponent microfabricated structures
US7056661B2 (en) 1999-05-19 2006-06-06 Cornell Research Foundation, Inc. Method for sequencing nucleic acid molecules
CA2377707A1 (en) 1999-06-22 2000-12-28 Invitrogen Corporation Improved primers and methods for the detection and discrimination of nucleic acids
US6929030B2 (en) 1999-06-28 2005-08-16 California Institute Of Technology Microfabricated elastomeric valve and pump systems
US6818395B1 (en) * 1999-06-28 2004-11-16 California Institute Of Technology Methods and apparatus for analyzing polynucleotide sequences
US6899137B2 (en) 1999-06-28 2005-05-31 California Institute Of Technology Microfabricated elastomeric valve and pump systems
CA2388528A1 (en) 1999-11-04 2001-05-10 California Institute Of Technology Methods and apparatus for analyzing polynucleotide sequences
DE60000109T2 (en) 1999-06-28 2002-11-07 California Inst Of Techn Elastomeric micropump and microvalve systems
US6533914B1 (en) * 1999-07-08 2003-03-18 Shaorong Liu Microfabricated injector and capillary array assembly for high-resolution and high throughput separation
US20040053290A1 (en) * 2000-01-11 2004-03-18 Terbrueggen Robert Henry Devices and methods for biochip multiplexing
CA2427669A1 (en) 2000-11-03 2002-06-06 Clinical Micro Sensors, Inc. Devices and methods for biochip multiplexing
WO2001007159A2 (en) * 1999-07-28 2001-02-01 Genset Integration of biochemical protocols in a continuous flow microfluidic device
US6977145B2 (en) 1999-07-28 2005-12-20 Serono Genetics Institute S.A. Method for carrying out a biochemical protocol in continuous flow in a microreactor
US6423536B1 (en) 1999-08-02 2002-07-23 Molecular Dynamics, Inc. Low volume chemical and biochemical reaction system
US6524456B1 (en) * 1999-08-12 2003-02-25 Ut-Battelle, Llc Microfluidic devices for the controlled manipulation of small volumes
US6824663B1 (en) 1999-08-27 2004-11-30 Aclara Biosciences, Inc. Efficient compound distribution in microfluidic devices
US6846626B1 (en) 1999-09-01 2005-01-25 Genome Technologies, Llc Method for amplifying sequences from unknown DNA
US6623945B1 (en) 1999-09-16 2003-09-23 Motorola, Inc. System and method for microwave cell lysing of small samples
EP1218543A2 (en) 1999-09-29 2002-07-03 Solexa Ltd. Polynucleotide sequencing
AU7747600A (en) 1999-10-01 2002-05-15 University Of California, The Microfabricated liquid sample loading system
JP3896739B2 (en) * 1999-10-29 2007-03-22 株式会社日立製作所 Capillary electrophoresis device
US6705345B1 (en) * 1999-11-08 2004-03-16 The Trustees Of Boston University Micro valve arrays for fluid flow control
US6120184A (en) 1999-11-17 2000-09-19 Stone Container Corporation Bag apparatus with reclosable pour spout
US6432290B1 (en) 1999-11-26 2002-08-13 The Governors Of The University Of Alberta Apparatus and method for trapping bead based reagents within microfluidic analysis systems
CA2290731A1 (en) 1999-11-26 2001-05-26 D. Jed Harrison Apparatus and method for trapping bead based reagents within microfluidic analysis system
JP4567935B2 (en) * 1999-12-27 2010-10-27 株式会社東芝 Hydrogen storage alloy, secondary battery, hybrid car and electric vehicle
CA2398107C (en) 2000-01-28 2013-11-19 Althea Technologies, Inc. Methods for analysis of gene expression
SE0000300D0 (en) * 2000-01-30 2000-01-30 Amersham Pharm Biotech Ab Microfluidic assembly, covering method for the manufacture of the assembly and the use of the assembly
US6807490B1 (en) 2000-02-15 2004-10-19 Mark W. Perlin Method for DNA mixture analysis
KR20020089357A (en) 2000-02-23 2002-11-29 자이오믹스, 인코포레이티드 Chips having elevated sample surfaces
IL151390A (en) 2000-02-28 2007-06-03 Adsil Lc Silane-based coating compositions, coated articles obtained therefrom and methods of using same
CA2402116A1 (en) 2000-03-07 2001-09-13 Foret Frantisek Parallel array of independent thermostats for column separations
WO2001072335A2 (en) * 2000-03-28 2001-10-04 Queen's University At Kingston Methods for effecting neuroprotection using a potassium channel modulator
SE0001768D0 (en) 2000-05-12 2000-05-12 Helen Andersson Microfluidic flow cell for the manipulation of particles
US6531282B1 (en) * 2000-05-30 2003-03-11 Oligotrail, Llc Multiplex amplification and analysis of selected STR loci
US7351376B1 (en) 2000-06-05 2008-04-01 California Institute Of Technology Integrated active flux microfluidic devices and methods
WO2002001081A2 (en) 2000-06-23 2002-01-03 Micronics, Inc. Valve for use in microfluidic structures
US6885982B2 (en) 2000-06-27 2005-04-26 Fluidigm Corporation Object oriented microfluidic design method and system
US6829753B2 (en) 2000-06-27 2004-12-07 Fluidigm Corporation Microfluidic design automation method and system
US6734401B2 (en) * 2000-06-28 2004-05-11 3M Innovative Properties Company Enhanced sample processing devices, systems and methods
US6531041B1 (en) * 2000-07-20 2003-03-11 Symyx Technologies, Inc. Multiplexed capillary electrophoresis system with rotatable photodetector
US7004184B2 (en) * 2000-07-24 2006-02-28 The Reagents Of The University Of Michigan Compositions and methods for liquid metering in microchannels
US20040013536A1 (en) * 2001-08-31 2004-01-22 Hower Robert W Micro-fluidic pump
EP1978110B1 (en) 2000-09-06 2010-05-26 Transnetyx, Inc. Computer-based method and system for screening genomic DNA
AU9087901A (en) 2000-09-15 2002-03-26 California Inst Of Techn Microfabricated crossflow devices and methods
EP1319716B1 (en) * 2000-09-22 2013-07-03 Hitachi, Ltd. Method of analyzing nucleic acid
US7097809B2 (en) 2000-10-03 2006-08-29 California Institute Of Technology Combinatorial synthesis system
US7258774B2 (en) 2000-10-03 2007-08-21 California Institute Of Technology Microfluidic devices and methods of use
US6782746B1 (en) 2000-10-24 2004-08-31 Sandia National Laboratories Mobile monolithic polymer elements for flow control in microfluidic devices
EP1202054B1 (en) 2000-10-25 2003-01-08 Bruker BioSpin GmbH LC-NMR system, comprising a device for trapping at least one component of a chromatography flow
US6871476B2 (en) 2000-11-13 2005-03-29 Stefan Tobolka Heat sealing and cutting mechanism and container forming apparatus incorporating the same
US6951632B2 (en) 2000-11-16 2005-10-04 Fluidigm Corporation Microfluidic devices for introducing and dispensing fluids from microfluidic systems
US6521188B1 (en) * 2000-11-22 2003-02-18 Industrial Technology Research Institute Microfluidic actuator
US6527003B1 (en) * 2000-11-22 2003-03-04 Industrial Technology Research Micro valve actuator
GB0028647D0 (en) 2000-11-24 2001-01-10 Nextgen Sciences Ltd Apparatus for chemical assays
SE0004351D0 (en) * 2000-11-27 2000-11-27 Helen Andersson System and method for time-controlled liquid handling of reactions and processes in a microfluidic flow cell system
US20020123538A1 (en) * 2000-12-29 2002-09-05 Peiguang Zhou Hot-melt adhesive based on blend of amorphous and crystalline polymers for multilayer bonding
EP1350029B1 (en) 2001-01-08 2014-09-10 President and Fellows of Harvard College Valves and pumps for microfluidic systems and method for making microfluidic systems
US20020098097A1 (en) * 2001-01-22 2002-07-25 Angad Singh Magnetically-actuated micropump
US7223363B2 (en) * 2001-03-09 2007-05-29 Biomicro Systems, Inc. Method and system for microfluidic interfacing to arrays
WO2002082057A2 (en) 2001-04-03 2002-10-17 Micronics, Inc. Split focusing cytometer
WO2002081729A2 (en) * 2001-04-06 2002-10-17 California Institute Of Technology Nucleic acid amplification utilizing microfluidic devices
WO2002081183A1 (en) * 2001-04-06 2002-10-17 Fluidigm Corporation Polymer surface modification
US6752922B2 (en) 2001-04-06 2004-06-22 Fluidigm Corporation Microfluidic chromatography
US6802342B2 (en) 2001-04-06 2004-10-12 Fluidigm Corporation Microfabricated fluidic circuit elements and applications
ES2287351T3 (en) 2001-04-25 2007-12-16 Cornell Research Foundation, Inc. Devices and methods for cell systems based on pharmacocinetics.
JP2002370200A (en) 2001-06-12 2002-12-24 Kawamura Inst Of Chem Res Method of manufacturing miniature valve mechanism
US6629820B2 (en) 2001-06-26 2003-10-07 Micralyne Inc. Microfluidic flow control device
GB0119959D0 (en) * 2001-08-16 2001-10-10 Sec Dep Of The Home Department Improvements in and relating to analysis
US20030095897A1 (en) * 2001-08-31 2003-05-22 Grate Jay W. Flow-controlled magnetic particle manipulation
US6852287B2 (en) * 2001-09-12 2005-02-08 Handylab, Inc. Microfluidic devices having a reduced number of input and output connections
JP3685119B2 (en) * 2001-10-18 2005-08-17 株式会社日立製作所 Biomolecule recovery method
JP4381670B2 (en) 2001-10-30 2009-12-09 株式会社日立製作所 Reactor
EP1442137A4 (en) * 2001-11-07 2005-08-31 Applera Corp Universal nucleotides for nucleic acid analysis
AU2002351187A1 (en) 2001-11-30 2003-06-17 Fluidigm Corporation Microfluidic device and methods of using same
US7691333B2 (en) 2001-11-30 2010-04-06 Fluidigm Corporation Microfluidic device and methods of using same
US6864480B2 (en) 2001-12-19 2005-03-08 Sau Lan Tang Staats Interface members and holders for microfluidic array devices
US6532997B1 (en) * 2001-12-28 2003-03-18 3M Innovative Properties Company Sample processing device with integral electrophoresis channels
US6581441B1 (en) 2002-02-01 2003-06-24 Perseptive Biosystems, Inc. Capillary column chromatography process and system
US6923907B2 (en) 2002-02-13 2005-08-02 Nanostream, Inc. Separation column devices and fabrication methods
US6685442B2 (en) * 2002-02-20 2004-02-03 Sandia National Laboratories Actuator device utilizing a conductive polymer gel
CA2478964A1 (en) * 2002-03-11 2003-09-25 Athenix Corporation Integrated system for high throughput capture of genetic diversity
US7312085B2 (en) 2002-04-01 2007-12-25 Fluidigm Corporation Microfluidic particle-analysis systems
EP2283917A3 (en) 2002-05-09 2012-08-01 The University of Chicago Device and method for pressure-driven plug transport and reaction
US20030217923A1 (en) * 2002-05-24 2003-11-27 Harrison D. Jed Apparatus and method for trapping bead based reagents within microfluidic analysis systems
US7215640B2 (en) * 2002-07-11 2007-05-08 Hitachi, Ltd. Method and apparatus for path configuration in networks
US20040101444A1 (en) 2002-07-15 2004-05-27 Xeotron Corporation Apparatus and method for fluid delivery to a hybridization station
US7943393B2 (en) * 2003-07-14 2011-05-17 Phynexus, Inc. Method and device for extracting an analyte
US6786708B2 (en) 2002-07-18 2004-09-07 The Regents Of The University Of Michigan Laminated devices and methods of making same
US20040018611A1 (en) 2002-07-23 2004-01-29 Ward Michael Dennis Microfluidic devices for high gradient magnetic separation
WO2004011681A1 (en) 2002-07-26 2004-02-05 Applera Corporation Microfluidic device including purification column with excess diluent, and method
US7198759B2 (en) 2002-07-26 2007-04-03 Applera Corporation Microfluidic devices, methods, and systems
US7244961B2 (en) 2002-08-02 2007-07-17 Silicon Valley Scientific Integrated system with modular microfluidic components
ES2282682T3 (en) * 2002-08-02 2007-10-16 Ge Healthcare (Sv) Corp. Integrated design of microchips.
US20040038385A1 (en) * 2002-08-26 2004-02-26 Langlois Richard G. System for autonomous monitoring of bioagents
US20040197845A1 (en) 2002-08-30 2004-10-07 Arjang Hassibi Methods and apparatus for pathogen detection, identification and/or quantification
DE60312186T2 (en) * 2002-09-06 2007-11-08 Epigem Ltd. Modular microfluid system
US7157228B2 (en) * 2002-09-09 2007-01-02 Bioarray Solutions Ltd. Genetic analysis and authentication
JP3725109B2 (en) 2002-09-19 2005-12-07 コニカミノルタホールディングス株式会社 Microfluidic device
TW590982B (en) 2002-09-27 2004-06-11 Agnitio Science & Technology I Micro-fluid driving device
WO2004040001A2 (en) * 2002-10-02 2004-05-13 California Institute Of Technology Microfluidic nucleic acid analysis
US7217542B2 (en) 2002-10-31 2007-05-15 Hewlett-Packard Development Company, L.P. Microfluidic system for analyzing nucleic acids
US20040086872A1 (en) 2002-10-31 2004-05-06 Childers Winthrop D. Microfluidic system for analysis of nucleic acids
JP2004180594A (en) 2002-12-04 2004-07-02 Shimadzu Corp Cell-culturing device
US20080047836A1 (en) * 2002-12-05 2008-02-28 David Strand Configurable Microfluidic Substrate Assembly
EP1579013A4 (en) 2002-12-13 2007-02-28 Gene Codes Forensics Inc Method for profiling and identifying persons by using data samples
US20050266582A1 (en) 2002-12-16 2005-12-01 Modlin Douglas N Microfluidic system with integrated permeable membrane
EP1594694A4 (en) 2002-12-30 2010-01-20 Univ California Methods and apparatus for pathogen detection and analysis
US7049558B2 (en) 2003-01-27 2006-05-23 Arcturas Bioscience, Inc. Apparatus and method for heating microfluidic volumes and moving fluids
US7575865B2 (en) 2003-01-29 2009-08-18 454 Life Sciences Corporation Methods of amplifying and sequencing nucleic acids
US7244567B2 (en) * 2003-01-29 2007-07-17 454 Life Sciences Corporation Double ended sequencing
WO2004067162A2 (en) 2003-01-30 2004-08-12 Ciphergen Biosystems Inc. Apparatus for microfluidic processing and reading of biochip arrays
US7338637B2 (en) 2003-01-31 2008-03-04 Hewlett-Packard Development Company, L.P. Microfluidic device with thin-film electronic devices
JP4119275B2 (en) 2003-02-18 2008-07-16 忠弘 大見 Diaphragm valve for vacuum exhaust system
US7476363B2 (en) 2003-04-03 2009-01-13 Fluidigm Corporation Microfluidic devices and methods of using same
WO2004092331A2 (en) 2003-04-08 2004-10-28 Li-Cor, Inc. Composition and method for nucleic acid sequencing
WO2004094020A2 (en) 2003-04-17 2004-11-04 Fluidigm Corporation Crystal growth devices and systems, and methods for using same
WO2004098757A2 (en) 2003-05-06 2004-11-18 New Jersey Institute Of Technology Microfluidic mixing using flow pulsing
FR2855076B1 (en) 2003-05-21 2006-09-08 Inst Curie Microfluidic device
US7063304B2 (en) 2003-07-11 2006-06-20 Entegris, Inc. Extended stroke valve and diaphragm
US7357898B2 (en) 2003-07-31 2008-04-15 Agency For Science, Technology And Research Microfluidics packages and methods of using same
US20050047967A1 (en) * 2003-09-03 2005-03-03 Industrial Technology Research Institute Microfluidic component providing multi-directional fluid movement
JP2007506430A (en) 2003-09-23 2007-03-22 ユニヴァーシティー オブ ミズーリ Polynucleotide synthesis method using thermostable enzyme
JPWO2005049196A1 (en) 2003-11-21 2007-12-27 株式会社荏原製作所 Microchip device using liquid
US7939249B2 (en) 2003-12-24 2011-05-10 3M Innovative Properties Company Methods for nucleic acid isolation and kits using a microfluidic device and concentration step
AT462493T (en) 2004-02-02 2010-04-15 Silicon Valley Scient Inc Integrated system with modular microfluidic components
US7407757B2 (en) * 2005-02-10 2008-08-05 Population Genetics Technologies Genetic analysis by sequence-specific sorting
US8043849B2 (en) 2004-02-24 2011-10-25 Thermal Gradient Thermal cycling device
JP5196422B2 (en) 2004-03-05 2013-05-15 ドゥンアン、マイクロスタック、インク Selective bonding for microvalve formation
ES2553097T3 (en) 2004-05-03 2015-12-04 Handylab, Inc. Processing of samples containing polynucleotides
JP4683538B2 (en) 2004-05-06 2011-05-18 セイコーインスツル株式会社 Analysis system and analysis method including microchip for analysis
JP3952036B2 (en) 2004-05-13 2007-08-01 コニカミノルタセンシング株式会社 Microfluidic device, test solution test method and test system
US7799553B2 (en) 2004-06-01 2010-09-21 The Regents Of The University Of California Microfabricated integrated DNA analysis system
US20050272144A1 (en) 2004-06-08 2005-12-08 Konica Minolta Medical & Graphic, Inc. Micro-reactor for improving efficiency of liquid mixing and reaction
US7585663B2 (en) 2004-08-26 2009-09-08 Applied Biosystems, Llc Thermal device, system, and method, for fluid processing device
WO2006032044A2 (en) 2004-09-15 2006-03-23 Microchip Biotechnologies, Inc. Microfluidic devices
US20060057209A1 (en) * 2004-09-16 2006-03-16 Predicant Biosciences, Inc. Methods, compositions and devices, including microfluidic devices, comprising coated hydrophobic surfaces
JP2006090386A (en) 2004-09-22 2006-04-06 Kitz Sct:Kk Diaphragm valve
GB0421529D0 (en) 2004-09-28 2004-10-27 Landegren Gene Technology Ab Microfluidic structure
WO2006044458A2 (en) 2004-10-13 2006-04-27 University Of Virginia Patent Foundation Electrostatic actuation for management of flow
US7832429B2 (en) * 2004-10-13 2010-11-16 Rheonix, Inc. Microfluidic pump and valve structures and fabrication methods
JP4850072B2 (en) * 2004-11-22 2012-01-11 日水製薬株式会社 Microchip
KR100634525B1 (en) 2004-11-23 2006-10-16 삼성전자주식회사 Microfluidic device comprising a microchannel disposed of a plurality of electromagnets, method for mixing a sample and method for lysis cells using the same
US8206593B2 (en) 2004-12-03 2012-06-26 Fluidigm Corporation Microfluidic chemical reaction circuits
CA2592204C (en) 2004-12-23 2013-03-12 I-Stat Corporation Nucleic acid diagnostics system and methods
US20060163143A1 (en) 2005-01-26 2006-07-27 Chirica Gabriela S Microliter scale solid phase extraction devices
CA2496481A1 (en) 2005-02-08 2006-08-09 Mds Inc., Doing Business Through It's Mds Sciex Division Method and apparatus for sample deposition
WO2006085443A1 (en) * 2005-02-10 2006-08-17 Matsushita Electric Industrial Co., Ltd. Fluid chip, control method for movement of fluid employing it, and chemical reactor
US20060210998A1 (en) * 2005-03-18 2006-09-21 Christiane Kettlitz Determination of antibiotic resistance in staphylococcus aureus
US20070017812A1 (en) * 2005-03-30 2007-01-25 Luc Bousse Optimized Sample Injection Structures in Microfluidic Separations
US20070015179A1 (en) 2005-04-26 2007-01-18 Trustees Of Boston University Plastic microfluidic chip and methods for isolation of nucleic acids from biological samples
US20060263789A1 (en) * 2005-05-19 2006-11-23 Robert Kincaid Unique identifiers for indicating properties associated with entities to which they are attached, and methods for using
US20070020654A1 (en) * 2005-05-19 2007-01-25 Affymetrix, Inc. Methods and kits for preparing nucleic acid samples
US8206974B2 (en) 2005-05-19 2012-06-26 Netbio, Inc. Ruggedized apparatus for analysis of nucleic acid and proteins
US7316766B2 (en) 2005-05-27 2008-01-08 Taidoc Technology Corporation Electrochemical biosensor strip
WO2007002579A2 (en) * 2005-06-23 2007-01-04 Bioveris Corporation Assay cartridges and methods for point of care instruments
US7817273B2 (en) 2005-06-30 2010-10-19 Life Technologies Corporation Two-dimensional spectral imaging system
US20070031865A1 (en) * 2005-07-07 2007-02-08 David Willoughby Novel Process for Construction of a DNA Library
US7428848B2 (en) * 2005-08-09 2008-09-30 Cfd Research Corporation Electrostatic sampler and method
CA2621632A1 (en) 2005-09-30 2007-04-12 Caliper Life Sciences, Inc. Microfluidic device for purifying a biological component using magnetic beads
US7709544B2 (en) 2005-10-25 2010-05-04 Massachusetts Institute Of Technology Microstructure synthesis by flow lithography and polymerization
US20080311585A1 (en) 2005-11-02 2008-12-18 Affymetrix, Inc. System and method for multiplex liquid handling
US20070113908A1 (en) 2005-11-18 2007-05-24 The Ohio State University And Bioloc, Inc. Valve for microfluidic chips
US20070122819A1 (en) 2005-11-25 2007-05-31 Industrial Technology Research Institute Analyte assay structure in microfluidic chip for quantitative analysis and method for using the same
US7763453B2 (en) 2005-11-30 2010-07-27 Micronics, Inc. Microfluidic mixing and analytic apparatus
JP4593451B2 (en) 2005-12-05 2010-12-08 セイコーインスツル株式会社 Microreactor system and liquid feeding method
US7976795B2 (en) 2006-01-19 2011-07-12 Rheonix, Inc. Microfluidic systems
CN101004423B (en) 2006-01-19 2011-12-28 博奥生物有限公司 Fluid sample analysis cartridge system
US7749365B2 (en) 2006-02-01 2010-07-06 IntegenX, Inc. Optimized sample injection structures in microfluidic separations
KR20080096567A (en) 2006-02-03 2008-10-30 마이크로칩 바이오테크놀로지스, 인크. Microfluidic devices
WO2007106580A2 (en) 2006-03-15 2007-09-20 Micronics, Inc. Rapid magnetic flow assays
US7766033B2 (en) 2006-03-22 2010-08-03 The Regents Of The University Of California Multiplexed latching valves for microfluidic devices and processors
US9063132B2 (en) * 2006-03-29 2015-06-23 Inverness Medical Switzerland Gmbh Assay device and method
KR100785010B1 (en) 2006-04-06 2007-12-11 삼성전자주식회사 Method and apparatus for the purification of nucleic acids on hydrophilic surface of solid support using hydrogen bonding
EP2481815B1 (en) * 2006-05-11 2016-01-27 Raindance Technologies, Inc. Microfluidic devices
WO2007136840A2 (en) * 2006-05-20 2007-11-29 Codon Devices, Inc. Nucleic acid library design and assembly
US8296088B2 (en) 2006-06-02 2012-10-23 Luminex Corporation Systems and methods for performing measurements of one or more materials
US8137912B2 (en) 2006-06-14 2012-03-20 The General Hospital Corporation Methods for the diagnosis of fetal abnormalities
EP2041573B1 (en) 2006-06-23 2019-09-04 PerkinElmer Health Sciences, Inc. Methods and devices for microfluidic point-of-care immunoassays
US7629124B2 (en) 2006-06-30 2009-12-08 Canon U.S. Life Sciences, Inc. Real-time PCR in micro-channels
CA2658533C (en) 2006-07-28 2016-03-15 Qiagen Gmbh Device for processing samples
WO2008024319A2 (en) 2006-08-20 2008-02-28 Codon Devices, Inc. Microfluidic devices for nucleic acid assembly
US20080131327A1 (en) 2006-09-28 2008-06-05 California Institute Of Technology System and method for interfacing with a microfluidic chip
WO2008052138A2 (en) * 2006-10-25 2008-05-02 The Regents Of The University Of California Inline-injection microdevice and microfabricated integrated dna analysis system using same
US20080242560A1 (en) 2006-11-21 2008-10-02 Gunderson Kevin L Methods for generating amplified nucleic acid arrays
EP3285067A1 (en) 2006-12-14 2018-02-21 Life Technologies Corporation Apparatus and methods for measuring analytes using fet arrays
US20080179255A1 (en) 2007-01-29 2008-07-31 Searete Llc, A Limited Liability Corporation Of The State Of Delaware Fluidic devices
EP2121983A2 (en) * 2007-02-02 2009-11-25 Illumina Cambridge Limited Methods for indexing samples and sequencing multiple nucleotide templates
EP2109666A4 (en) 2007-02-05 2011-09-14 Integenx Inc Microfluidic and nanofluidic devices, systems, and applications
US8877518B2 (en) * 2007-02-06 2014-11-04 The Trustees Of The University Of Pennsylvania Multiplexed nanoscale electrochemical sensors for multi-analyte detection
CN101663089A (en) 2007-04-04 2010-03-03 微点生物技术有限公司 micromachined electrowetting microfluidic valve
KR20100021565A (en) * 2007-04-04 2010-02-25 네트워크 바이오시스템즈, 인코퍼레이티드 Methods for rapid multiplexed amplification of target nucleic acids
US8702976B2 (en) 2007-04-18 2014-04-22 Ondavia, Inc. Hand-held microfluidic testing device
WO2008147530A1 (en) 2007-05-24 2008-12-04 The Regents Of The University Of California Integrated fluidics devices with magnetic sorting
US20100111770A1 (en) 2007-06-07 2010-05-06 Samsung Electronics Co., Ltd. Microfluidic Chip and Method of Fabricating The Same
EP2167216B1 (en) 2007-06-29 2012-05-02 The President and Fellows of Harvard College Density-based methods for separation of materials, monitoring of solid supported reactions and measuring densities of small liquid volumes and solids
WO2009008236A1 (en) 2007-07-10 2009-01-15 Konica Minolta Medical & Graphic, Inc. Method of mixing liquids in micro-inspection chip and inspection instrument
US9618139B2 (en) 2007-07-13 2017-04-11 Handylab, Inc. Integrated heater and magnetic separator
US8454906B2 (en) 2007-07-24 2013-06-04 The Regents Of The University Of California Microfabricated droplet generator for single molecule/cell genetic analysis in engineered monodispersed emulsions
DE102007035721B4 (en) 2007-07-30 2019-02-07 Robert Bosch Gmbh Microvalve, method of manufacturing a microvalve and micropump
WO2009020435A1 (en) 2007-08-07 2009-02-12 Agency For Science, Technology And Research Integrated microfluidic device for gene synthesis
JP2010536565A (en) * 2007-08-23 2010-12-02 シンベニオ・バイオシステムズ・インコーポレーテッドCynvenio Biosystems Incorporated Magnetic sorting system for traps for target species
EP2214823A1 (en) * 2007-11-26 2010-08-11 Atonomics A/S Integrated separation and detection cartridge with means and method for increasing signal to noise ratio
CN101990516B (en) 2008-01-22 2015-09-09 英特基因有限公司 Multiplex sample preparation system and the use in integrated analysis system thereof
WO2009117611A2 (en) 2008-03-19 2009-09-24 Cynvenio Biosystems, Llc Trapping magnetic cell sorting system
WO2009129415A1 (en) 2008-04-16 2009-10-22 Cynvenio Biosystems, Llc Magnetic separation system with pre and post processing modules
US7867713B2 (en) * 2008-04-21 2011-01-11 Lawrence Livermore National Security, Llc Polymerase chain reaction system using magnetic beads for analyzing a sample that includes nucleic acid
US20090269504A1 (en) 2008-04-24 2009-10-29 Momentive Performance Materials Inc. Flexible hardcoats and substrates coated therewith
KR100960066B1 (en) 2008-05-14 2010-05-31 삼성전자주식회사 Microfluidic device containing lyophilized reagent therein and analysing method using the same
US8323568B2 (en) 2008-06-13 2012-12-04 Honeywell International Inc. Magnetic bead assisted sample conditioning system
US8092761B2 (en) 2008-06-20 2012-01-10 Silverbrook Research Pty Ltd Mechanically-actuated microfluidic diaphragm valve
US7976789B2 (en) 2008-07-22 2011-07-12 The Board Of Trustees Of The University Of Illinois Microfluidic device for preparing mixtures
US8617488B2 (en) 2008-08-07 2013-12-31 Fluidigm Corporation Microfluidic mixing and reaction systems for high efficiency screening
JP2012501658A (en) 2008-09-05 2012-01-26 ライフ テクノロジーズ コーポレーション Methods and systems for nucleic acid sequencing validation, calibration, and standardization
JP5311518B2 (en) 2008-10-06 2013-10-09 コーニンクレッカ フィリップス エヌ ヴェ Microfluidic device
US9422409B2 (en) 2008-10-10 2016-08-23 Massachusetts Institute Of Technology Method of hydrolytically stable bonding of elastomers to substrates
EP2350652A2 (en) 2008-10-10 2011-08-03 Cnrs-Dae Cell sorting device
US20100233696A1 (en) 2008-11-04 2010-09-16 Helicos Biosciences Corporation Methods, flow cells and systems for single cell analysis
WO2010077322A1 (en) 2008-12-31 2010-07-08 Microchip Biotechnologies, Inc. Instrument with microfluidic chip
NZ620826A (en) 2009-02-03 2015-09-25 Netbio Inc Nucleic acid purification
US20100221726A1 (en) 2009-02-09 2010-09-02 Frederic Zenhausern Relating to devices
RU2011142784A (en) 2009-03-23 2013-04-27 Конинклейке Филипс Электроникс Н.В. Manipulation by magnetic particles in a biological sample
US20100243916A1 (en) 2009-03-30 2010-09-30 Lockheed Martin Corporation Modular optical diagnostic platform for chemical and biological target diagnosis and detection
US20110003301A1 (en) * 2009-05-08 2011-01-06 Life Technologies Corporation Methods for detecting genetic variations in dna samples
WO2010141326A1 (en) 2009-06-02 2010-12-09 Integenx Inc. Fluidic devices with diaphragm valves
US20110065101A1 (en) 2009-06-04 2011-03-17 Lockheed Martin Corporation Multiple-sample microfluidic chip for DNA analysis
SG176669A1 (en) 2009-06-05 2012-01-30 Integenx Inc Universal sample preparation system and use in an integrated analysis system
WO2011003941A1 (en) 2009-07-07 2011-01-13 Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. Device for detection of biological agents
US8551787B2 (en) 2009-07-23 2013-10-08 Fluidigm Corporation Microfluidic devices and methods for binary mixing
GB2472236A (en) 2009-07-29 2011-02-02 Iti Scotland Ltd Apparatus for analysing microfluidic devices
EP2480324A4 (en) 2009-09-21 2013-10-02 Akonni Biosystems Magnetic lysis method and device
GB2473868A (en) 2009-09-28 2011-03-30 Invitrogen Dynal As Apparatus and method of automated processing of biological samples
TWI421495B (en) 2009-10-30 2014-01-01 Univ Nat Cheng Kung Microfluidic chip
WO2011084703A2 (en) 2009-12-21 2011-07-14 Advanced Liquid Logic, Inc. Enzyme assays on a droplet actuator
US8580505B2 (en) 2010-02-26 2013-11-12 Life Technologies Corporation Fast PCR for STR genotyping

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011133014A1 (en) * 2010-04-19 2011-10-27 Mimos Berhad Planar micropump with integrated microvalves
US8778282B2 (en) 2010-06-15 2014-07-15 Samsung Electronics Co., Ltd. Microfluidic device having microvalve
WO2012144794A2 (en) * 2011-04-19 2012-10-26 주식회사 바이오포커스 Method for manufacturing a microvalve device mounted on a lab-on-a-chip, and microvalve device manufactured by same
WO2012144794A3 (en) * 2011-04-19 2013-01-17 주식회사 바이오포커스 Method for manufacturing a microvalve device mounted on a lab-on-a-chip, and microvalve device manufactured by same
KR101299438B1 (en) * 2011-04-19 2013-08-29 손문탁 method for manufacturing microvalve element mounted on lab-on-a-chip, and microvalve element produced thereby
KR101404657B1 (en) * 2013-04-26 2014-06-09 주식회사 바이오록스 Manifold Package for Driving Lab-on-a-Chip That Can Supply Vacuum, Air Vent and Compressed Air

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